Biology 2

Chapter 18 NOTES

Eukaryotic gene regulation is more complex than in prokaryotes, because eukaryotes:

· Have larger, more complex genomes. This requires that eukaryotic DNA be more complexly organized than prokaryotic DNA.

· Require cell specialization or differentiation.

1. Each cell of a multicellular eukaryote expresses only a small fraction of its genome

Cellular differentiation = Divergence in structure and function of different cell types, as they become specialized during an organism’s development

· Usually, it is DNA transcription that is controlled.

· Eukaryotes have more complex chromosomal structure, gene organization and cell structure than prokaryotes, which offer added opportunities for controlling gene expression.

II. The structural organization of chromatin sets coarse controls one gene expression

Prokaryotic and eukaryotic cells both contain double-stranded DNA, but their genomes are organized differently.

Prokaryotic DNA is:

· Usually circular.

· Much smaller than eukaryotic DNA; it makes up a small nucleoid regions only visible with an electron microscope.

· Associated with only a few protein molecules.

· Less elaborately structured and folded than eukaryotic DNA; bacterial chromosomes have some additional structure as the DNA-protein fiber forms loops that are anchored to the plasma membrane.

Eukaryotic DNA is:

· Complexed with a large amount of protein to form chromatin.

· Highly extended and tangled during interphase.

· Condensed into short, thick, discrete chromosomes during mitosis; when stained, chromosomes are clearly visible with a light microscope.

Eukaryotic chromosomes contain an enormous amount of DNA, which requires an elaborate system of DNA packing to fit all of the cell’s DNA into the nucleus.

B. Nucleosomes, or “Beads on a String”

Histone proteins associated with DNA are responsible for the first level of DNA packing in eukaryotes.

Histones = Small proteins that are rich in basic amino acids and that bind to DNA, forming chromatin.

· Contain a high proportion of positively charged amino acids (arginine and lysine), which bind tightly to the negatively charged DNA.

· Are present in approximately equal amounts to DNA in eukaryotic ells.

· Are similar from one eukaryote to another, suggesting that histone genes have been highly conserved during evolution. There are five types of histones in eukaryotes.

Partially unfolded chromatin (DNA and its associated proteins) resembles beads spaced along the DNA string. Each beadlike structure is a nucleosome.

Nucleosome = The basic unit of DNA packing; it is formed form DNA wound around a protein core that consists of two copies each of four types of histone (H2A, H2B, H3, H4). A fifth histone (H1) may be present on DNA next to the nucleosome.

· Nucleosomes may control gene expression by controlling access of transcription proteins of DNA.

· Nucleosome heterogeneity may also help control gene expression; nucleosomes may differ in the extent of amino acid modification and in the type of nonhistone proteins present.

B. Higher Levels of DNA Packing

The 30-nm chromatin fiber is the next level of DNA packing.

· This structure consists of a tightly would coil with six nucleosomes per turn.

· Molecules of histone H1 pull the nucleosomes into a cylinder 30nm in diameter.

In the next level of higher-order packing, the 30-nm chromatin fiber forms looped domains, which:

· Are attached to a nonhistone protein scaffold.

· Contain 20,000 to 100,000 base pairs.

· Coil and fold, further compacting the chromatin into a mitotic chromosome characteristic of metaphase.

Interphase chromatin is much less condensed than mitotic chromatin, but it still exhibits higher-order packing.

· Its nucleosome string is usually coiled into a 30-nm fiber, which is folded into looped domains.

· Interphase looped domains attach to a scaffolding inside the nuclear envelope (nuclear lamina); this helps organize areas of active transcription.

· Chromatin fibers of different chromosomes do not become entangled as they eccupy-restricted areas within the nucleus.

Portions of some chromosomes remain highly condensed throughout the cell cycle, even during interphase. Such heterochromatin is not transcribed.

Heterochromatin = Chromatin that remains highly condensed during interphase and that is not actively transcribed.

Euchromatin = Chromatin that is less condensed during interphase and is actively transcribed; euchromatin becomes highly condensed during mitosis.

What is the function of heterochromatin in interphase cells?

· Since most heterochromatin is not transcribed, it may be a coarse control of gene expression.

· For example, Barr bodies in mammalian cells are X chromosomes that are mostly condensed into heterochromatin. In female somatic cells, one X chromosome is a Barr body, so the other X chromosome is the only one transcribed.

III. Noncoding sequences and gene duplication account for much of a eukaryotic genome

DNA in eukaryotic genomes is organized differently from that in prokaryotes.

· In prokaryotes, most DNA codes for protein (mRNA), tRNA or rRNA, and coding sequences are uninterrupted. Small amounts f noncoding DNA consist mainly of control sequences, such as promoters.

· In eukaryotes, most DNA does not encode protein or RNA, and coding sequences may be interrupted by long stretches of noncoding DNA (introns). Certain DNA sequence may be present in multiple copies.

C. Repetitive Sequences

About 10-25% of total DNA in higher eukaryotes is satellite DNA that consists f short (5 to 10 nucleotides) sequences that are tandemly repeated thousands of times.

Satellite DNA = In eukaryotic chromosomes, highly repetitive DNA consisting of short unusual nucleotide sequences that are tandemly repeated thousands of times.

· Called satellite DNA because its unusual nucleotide ratio gives it a density different from the rest of the cell’s DNA. Thus, during ultracentrifugation, satellite DNA separates out in a cesium chloride gradient as a “Satellite” band separate from the rest of the DNA.

· Is not transcribed and its function is not known. Since most satellite DNA in chromosomes is located at the tip and the centromere, scientists speculate that it plays a structural role during chromosome replication and chromatid separation in mitosis and meiosis.

It is known that short tandem repeats called telomeres - at the ends of eukaryotic chromosomes -are important in maintaining the integrity of the lagging DNA strand during replication.

Telomere = Series of short tandem repeats at the ends of eukaryotic chromosomes; prevents chromosomes from shortening with each replication cycle.

· Before an Okazaki fragment of the lagging DNA strand can be synthesized, RNA primers must be produced on a DNA template ahead of the sequence to be replicated

· Since such a template is not possible for the end of a linear DNA molecule, there must be a mechanism to prevent DNA strands from becoming shorter with each replication cycle.

· This end-replication problem is solved by the presence of special repeating telomeric sequences on the ends of linear chromosomes.

· To compensate for the loss of telomeric nucleotides that occurs each replication cycle, the enzyme telomerase periodically restores this repetitive sequence to the ends of DNA molecules.

· Telomeric sequences are similar among many organisms and contain a block of G nucleotides. For example, human chromosomes have 250-1500 repetitions of the base sequence TTAGGG (AATCCC on the complementary strand).

There are other highly repetitive sequences in eukaryotic genomes. For example,

· Some are tranposons; generally regarded as nonfunctional, they are associated with some diseases (e.g. neurofibromatosis-1 or elephant man’s disease and some cancers).

· Mutations can extend the repetitive sequences normally found within the boundary of genes and cause them to malfunction (e.g. fragile X syndrome an Huntington’s disease.)

·

D. Multigene Families

Most eukaryotic genes are unique sequences present as single copies in the genome, but some genes are part of a multigene family.

Multigene family = A collection of genes that are similar or identical in sequence and presumably of common ancestral origin; such genes may be clustered or dispersed in the genome.

Families of identical genes:

Probably arise from a single ancestral gene that has undergone repeated duplication. Such tandem gene duplication results from mistakes made during DNA replication and recombination.

Are usually clustered and almost exclusively genes for RNA products. (One exception is the gene family coding for histone proteins.)

Include genes for the major rRNA molecules; huge tandem repeats of these genes enable cells to make millions of ribosomes during active protein synthesis.

Families of nonidentical genes:

· Arise over time from mutations that accumulate in duplicated genes.

· Can be clustered on the same chromosome or scattered throughout the genome. (Note that, for carious reasons, gene sequences in tandem arrays on the same chromosome tend to stay very similar to one another. Transposition events that translocate variants of duplicated genes to different chromosomes help stabilize their differences and thus promote diversity.)

· May include pseudogenes or nonfunctional versions of the duplicated gene.

Pseudogene= Nonfunctional gene that has a DNA sequence similar to a functional gene; but, as a consequence of mutation, lacks sites necessary for expression.

A good example of how multigene families can evolve from a single ancestral gene is the globin gene family - actually two related families of genes that encode globins, the a and b polypeptide subunits of hemoglobin. Based on amino acid homologies, the evolutionary history has been reconstructed as follows:

Ancestral globin gene

Single –chain globin

Duplication

a b

Mutation

Transposition:

a b

a&b genes separate

z a e g b

Duplication and

mutations

z

· The original a andb genes evolved from duplication of a common ancestral globin gene. Gene duplication was followed by mutation.

· Transposition separated the a globin and b globin families, so they exist on different chromosomes.

· Subsequent episodes of geneduplication and mutation resulted in new genes and pseudogenes in each family.

The consequence is that each globin gene family consists of a group of similar, but not identical genes clustered on a chromosome.

· In each gene family, the genes are arranged in order of their expression. During development, genes are turned on or off in response to the organism’s changing environment as it develops from an embryo into a fetus, and then into an adult.

· At all times during development, functional hemoglobin consists of two a-like and two b-like polypeptides.

· During embryongenesis, the z and e forms predominate.

· About 10 weeks after fertilization, the products of the a-globin genes replace that of the z gene, and the g-chain gene products - g^G and the g^A globins - become more prevalent.

· Prior to birth, the b-gene product begins to replace the g^G and g^A globins, so by six months of age, the adult b-like globins, d and b, are present.

· In humans, embryonic and fetal hemoglobins have a higher affinity for oxygen than the adult forms, allowing efficient oxygen exchange between mother and developing fetus.

IV. The control of gene expression can occur at any step in the pathway from gene to functional protein

Chromatin organization:

· Packages DNA into a compact form that can be contained by the cell’s nucleus.

· Controls which DNA regions are available for transcription.

· Condensed heterochromatin is not expressed.

· A gene’s location relative to nucleosomes and to scaffold attachment sites influences ts expression.

Availability for transcription is only the first level of control of gene expression in eukaryotic cells. Additional levels of control allow cells to alter gene expression in response to changing conditions and needs.

Complexities in chromosome structure gene organization and cell structure provide opportunities fro the control of gene expression in eukaryotic cells. Th e steps of gene expression where gene regulation can occur are outlined below.

Nucleus

Chromatin

Pretranscriptional

Control

(Gene level)

Gene available for expression

Transcriptional

Control

Initial RNA transcript

MRNA transcript

CYTOPLASM

mRNA in cytoplasm

Posttranscriptional

Control

Polypeptide product

Functional protein

Protein in use

A. Organization of a typical Eukaryotic Gene: A review

Eukaryotic genes:

Contain introns, noncoding sequences that intervene within the coding sequence.

Contain a promoter sequence at the 5’ upstream end; RNA polymerase attaches to a promoter sequence and transcribes introns along with the coding sequences, or exons.

May be regulated by enhancers, other noncoding control sequences that can be located thousands of base pairs away from the promoter.

Enhancer = Noncoding DNA control sequence that enhances a gene’s transcription and tha is located thousands of bases away from the gene’s promoter.

The primary RNA transcript ( hnRNA) is processed into mature mRNA by:

Removal of introns.

Addition of a modified guanosine triphosphate cap at the 5’ end.

Addition of a poly-A tail at the 3’ end.

B. Transcriptional Control

In both prokaryotes and eukaryotes, transcription requires that RNA polymerase recognize and bind to DNA at the promoter. However, transcription in eukaryotes differs in two important ways, offering additional opportunities for transcriptional control:

Eukaryotic RNA polymerase cannot recognize the promoter without the help of general transcription factors - proteins that must assemble on DNA at the promoter before transcription can begin.

For example, recall from chapter 16 that most eukaryotic genes are transcribed by RNA polymerase II, which cannot recognize a promoter unless a specific general transcription factor is bond to the TATA box.

Gene regulatory proteins can influence the rate of transcription by speeding up or slowing down this assembly process at the promoter.

· Enhancer sequences, noncoding sequences thousands f nucleotide pairs away from the promoter, influence gene expression in eukaryotes.

Enhancer sequence = Noncoding DNA control sequence to which transcription factors (gene regulatory proteins) bind , controlling transcription of a structural gene; may be thousands of nucleotide pairs away from the promoter of the gene it influences.

Associations between transcription factors and enhancer sites are important transcriptional controls in eukaryotes. How do enhancers stimulate transcription of specific genes?

· One hypothesis is that a hairpin loop forms in DNA, bringing the transcription factor bound to enhancer into contact with transcription factors and polymerase at the promoter.

· Diverse ehancer sequences and their transcription factors may selectively activate gene expression at appropriate stag3es in cell development.

Control of transcription thus depends on selective bidning of gene regulatory proteisn to specific DNA sequences and proteins. This highly selective binding depends on molecular shape.

· There must by a complementary fit between the surfaces of a transcription factor and its specific DNA-binding site.

· Hundreds of transcription fators have been discovered; and though each of these proteins is unique, many recognize their DNA-binding sites with ony one of a few possible structural motifs of domains containing a helices or b sheets.

C. Posttranscriptiona Control

Transcription produces a primary transcript, but gene expression – the production of protein, tRNA, or rRNA – may be stopped or enhanced at any posttranscriptional step.

1. RNA processing and Export

Because eukaryotic cells have a nuclear envelope, translation is segregated from transcription. This offers additional opportunities for controlling gene expression.

Before leaving the nucleus, the initial mRNA transcript (pre-mRNA) is processed in two ways:

Both ends are covalently altered; a 5’ cap and a poly-A tail are added.

Introns are removed and the remaining exons are spliced together.

The processed mRNA transcript moves to the cytoplasm through pores in the nuclear envelope.

Once in the cytoplasm, the mRNA interacts with specific proteins and may associate with ribosomes to begin translation.

Little is known about how these posttranscriptional steps are regulated.

2. Regulation of mRNA Degredation

Protein synthesis is also controlled by mRNA’s lifespan in the cytoplasm.

Ø Prokarytotic mRNA molecules are degraded by enzymes after only a few minutes. Thus, bacteria can quickly alter patterns of protein synthesis in response to environmental change.

Ø Eukryotic mRNA molecules can exist for several hours or even weeks.

Ø The longevity of a mRNA molecules can exist for several hours or even weeks.

Ø The longevity of a mRNA affects how much protein synthesis it directs. Those that are viable longer can produce more of their protein.

Ø For example, long-lived mRNAs for hemoglobin are repeatedly translated in developing vertebrate red blood cells.

In some cases, mRNA molecules accumulate and a mass translation occurs after some control signal is received.

For example, prior to fertilization, the ovum produces and stores inactive mRNA that will be used later in development.

In cases such as these, the stored mRNA must not only be blocked form translation, but must be protected from degradation as well. mRNA degradation can be prevented by:

Rna-binding proteins. Some mRNA-binding proteins that repress translation, also prevent mRNA degradation.

Selective poly-A addition or removal. There is a minimum poly-A tail length required for mRNA stability and a longer length required for translation.

D. Translational and Posttranslational Control

Gene expression can also be regulated by mechanisms that control translation of mRNA into protein, most of these translational controls repress initiation; for example.

Binding of translation repressor protein to the 5’ and of a particular mRNA can prevent ribosome attachment.

Prior to fertilization, the ovum produces and stores inactive mRNA to be used later during the first embryonic cleavage.

The inactive mRNA is stored in the ovum’s cytosol until fertilization, when the sudden activation of an initiation factor triggers translation.

Delayed translation of stockpiled mRNA allows cells to respond quickly with a burst of protein synthesis when it si needed.

Posttranlational control is the last level of control for regulating gene expression.

Many eukaryotic polypeptides must be modified or transported before becoming biologically active. Such modifications include:

Cleavage of the polypeptide into smaller molecules (e.g. insulin).

Adding chemical groups, such as sugars.

Dispatching proteins targeted by signal sequences for specific sites.

Selective degradation of particular proteins and regulation of enzyme activity are also control mechanisms of gene expression.

E. The arrangement of Coordinately Controlled Genes

Coordinately controlled genes are arranged differently in a eukaryotic chromosome than in prokaryotic genomes.

Prokaryotic genes that are turned on and off together are foten clustered into operons; these adjacent genes share regulatory sites located at one end of the cluster. All genes of the operon are transcribed into one mRNA molecule and are translated together.

Eukarytoci genes coding for enzymes of a metabolic pathway are often scattered over different chromosomes. Even functionally related genes on the same chromosome have their own promoters and arre individually transcribed.

Eukaryotic genes can be coordinately expressed, even though they may be scattered throughout the genome.

Coordinatlely controlled genes are each associated with specific regulatory DNA sequences of enhancers. These sequences are recognized by a single type of transcription factor that activated or represses a group of genes in synchrony.

Examples of coordinate gene expression in eukaryotes includes:

Heat shock response. Exposure to high temperature activates genes coding for heat shock proteins, which help stabilize and repair heat-denatured proteins in the cell.

Steroid hormone action. Steroids activate protein receptors , and the protein receptor complex, in turn, activates genes. In a secondary response, proteins produced this way can activate Anita group of genes.

Cellular differentiation. During cellular differentiation, coordinately controlled genes producing particular sets of proteins are switched on and off.

E. Gene Expression and Differentiation

In multicellular organisms, cells differentiate into specialized cell types which become more organized into tissues and are characterized by tissue-specific gene expression.

Development fate of embryonic cells is determined by cytoplasmic content and cell position within the embryo; these factors influence what developmental chemical signals will be present.

Affecting only certain cells, these chemical signals activate transcription factors, which result in gene expression for other regulatory proteins.

Cascade of regulatory proteins, in turn, cause batteries of tissue-specific genes to be expressed.

V. Chemical signals that help control gene expression include hormones

Small molecules in the cell’s external environment can influence gene expression. Animal steroid hormones, for example, help control transcription by binding to regulatory proteins, altering their conformation.

As chemical messengers, hormones such as steroids are produced in one part of the body and then trace in the bloodstream to affect target organs else3where, far removed from the site of production.

The effect of steroids on gene expression in target cells was recognized first in insects.

A. Chromosome Puffs: Evidence for the Regulatory Role of Steroid Hormones in Insects

Evidence for transcriptional control and the influence of steroids on gene expression have come from studies of polytene chromosomes from the salivary gland and other tissues of insects.

Polytene chromosomes are single giant chromosomes produced by repeated DNA replication without cell division; rather than separating out as individual chromosomes, the hundreds of homologous chromosome copies remain next to each other.

· Polytene chromosomes are single giant chromosomes produced by a repeated DNA replication without cell division; rather than seperating out as individual chromosomes, the hundreds of homologous chromosome copies remain next to each other.

· At particular points of larval development, chromosome puffs - loops of decondensed DNA - appear at specific sites on the polytene chromosomes.

· Growing RNA transcripts detected by autoradiography, indicate that intense transcription occurs at puffs.

As a larva develops, puff location along the chromosome changes.

· At molting, some puffs disappear and new puffs form at other sites.

· Shifting puffs indicate that selective switching on and off of specific genes occurs during development.

· Ecdysone (molting hormone) can induce changes in puff patterns, demonstrating that gene regulation responds to specific chemical signals.

B. The action of Steroid Hormones in Vertebrates

Steroid hormones are chemical signals that can activate gene expression in target cells of vertebrates. The key steps of steroid hormone action are outlined below.

· Since steroids are lipid soluble, they readily diffuse across the plasma membrane into the cytoplasm.

· From the cytoplasm, the steroid enters the nucleus , where it binds to a soluble, steroid-receptor protein - a DNA-binding protein that can activate transcription of a particular gene. (Some steroids bind to receptors in the cytoplasm, and the hormone receptor complex moves as a unit into the nucleus.)

· In absence of steroid, an inhibitory protein binds to the steroid receptor and blocks its DNA-binding domain, preventing the receptor form binding to DNA.

· When steroid is present, its binding to the receptor causes release of inhibitory protein and activates the steroid receptor, so it can attach to DNA at enhancer sequences that control steroid-responsive genes.

Summary: The Control of Gene Expression in Eukaryotes

· Various cell types of multicellular organisms express different genes.

· Chromatin organization makes certain genes available for expression and other genes unavailable.

· Opportunities for the control of gene expression exist at each step in the pathway from gene to functional protein.

· Transcriptional control is important in determining which genes are expressed; selective binding of regulatory proteins to enhancer sequences in DNA, stimulates transcription of specific genes.

· Regulatory activity of some DNA-binding proteins is affected by certain hormones and other chemical cues.

VI. Chemical modification or relocation of DNA within a genome can alter gene expression

An organism’s genome is plastic, or changeable, in ways that attect the acailability of specific genes for expression.

· Gene may be available for expression in some cells and not others, or at some time in the organism’s development and not others.

· TGenes may, under some conditions, be amplified or made more available than usual.

· Changes in the physical arrangement of DNA, such as levels of DNA packing, affect gene expression. For example, genes in heterochramatin and mitotic chromosomes are not expressed.

The structural organization of an organism’s genome is also somewhat plastic; movement of DNA within the genome chemical modification of DNA influence gene expression.

A. Gene Amplification and Selective Gene Loss

Gene amplification may temporarily increase the number of gene copies at certain times in development.

Gene amplification = Selective synthesis of DNA, which results in multiple copies of a single gene. For example,

· Amphibian rRNA genes are selectively amplified in the ocyte, which:

· Results in a million or more additional copies of the rRNA genes that exist as extrachromosomal circle of DNA in the nucleoli.

· Permits the oocyte to make huge numbers of ribosomes that will produce the vast amounts of proteins needed when the egg is fertilized.

· Gene amplification occurs in cancer cells exposed to high concentrations of chemotherapuetic drugs.

· Some cancer cells survive chemotherapy, because they contain amplified genes conferring drug resistance.

· Increases drug resistance can be created experimentally by exposing a cell population to increasing drug doses and artificially selecting for surviving cells that have amplified drug-resistance genes.

Genes may also be selectively lost in certain tissues by elimination of chromosomes.

Chromosome diminution = Elimination of whole chromosomes or parts of chromosomes from certain cells early in embryonic development.

· For example, chromosome diminution occurs in gall midges during early development; all but two cells lost 32 of their 40 chromosomes during the first mitotic division after the 16-cell stage.

· The two cells that retain the complete genome are germ cells that will produce gametes in the adult. The other 14 cells become somatic cells with only 8 chromosomes.

B. Rearrangements in the Genome

Rearrangements of DNA in somatic cells can activate of inactivate specific genes.

1. Transposons

All organisms probably have Transposons that move DNA from one location to another within the genome. Transposons can rearrange the genome by:

· Inserting into the middle of a coding sequence of another gene; it can prevent the interrupted gene from functioning normally

· Inserting within a sequence that regulates transcription; the transposition may increase or decrease a protein’s production.

· Inserting its own gene just downstream form an active promoter that activates its transcription.

2. Immunoglobulin Genes

During cellular differentiation in mammals, permanent rearrangements of DNA segments occur in those genes that encode antibodies, or immunoglobulins.

Immunoglobulins = A class of proteins (antibodies) produced by B lymphocytes that specifically recognize and help combat viruses, bacteria, and other invaders of the body. Immunoglobulin molecules consist of:

· Four polypeptide chains held together by disulfide bridges.

· Each chain has two major parts:

· A constant region, which is the same for all antibodies of a particular class.

· A variable region, which gives an antibody the ability to recognize and bind to a specific foreign molecule.

B lymphocytes, which produce immunoglobulins, are a type of white blood cell found in the mammalian immune system.

· The human immune system contains millions of subpopulation of B lymphocytes that produce different antibodies.

· B lymphocytes are very specialized; each differential cell and its descendants produce only one specific antibody.

Antibody specificity and diversity are properties that emerge from the unique organization of the organization of the antibody gene, which is formed by a rearrangements of the genome during B

cell development.

· As an unspecialized cell differentiates into a B lymphocyte, its antibody gene is pieced together randomly from several DNA segments that are physically separated in the genome.

· In the genome of an embryonic cell, there is an intervening DNA sequence between the sequence coding for an antibody’s constant region and the site containing hundreds of coding sequences for the variable regions.

· As a B cell differentiates, the intervening DNA is deleted, and the DNA sequence for a variable region connects with the DNA sequence for a constant region, forming a continuos nucleotide sequence that will be transcribed.

· The primary RNA transcript is processed to form mRNA that is translated into one of the polypeptides of an antibody molecule.

· Antibody variation results from:

· Different combinations of variable and constant regions in the polypeptides.

· Different combination of polypeptides

C. DNA Methylation

DNA methylation = The addition of methyl groups (-CH₃) to bases of DNA, after DNA synthesis.

· Most plant and animal DNA contains methylate bases (usually cytosine); about 5% of the cytosine residues are methylated.

· May be a cellular mechanism for long-term control of gene expression. When researchers examine the same genes from different types of cells, they find:

· Genes that are not expressed (e.g. Barr bodies) are more heavily methylated than those that are expressed.

· Drugs that inhibit methylation can induce gene reactivation, even in Barr bodies.

· In vertebrate, DNA methylation reinforces developmental decisions made earlier by other mechanisms.

· For example, genes must be selectively turned on or off for normal cell differentiation to occur. DNA methylation insures that once a gene is turned off, it stays off.

· DNA methylation patterns are inherited and thus perpetuated as cells divide; clones of a cell lineage forming specialization tissues have a chemical record of regulatory events that occurred during early development.

VII. Cancer can result from the abnormal expression of genes that regulate cell growth and division

Cancer is a variety of diseases in which cells escape from the normal controls on growth and division, and it can result from mutations that alter normal gene expression in somatic cells. These mutations:

· Can be random and spontaneous

· Most likely occur as a result of environmental influences, such as:

· Infection by certain viruses.

· Exposure to carcinogens.

Carcinogens = Physical agents such as x-rays and chemical agents that cause cancer by mutating DNA.

Whether cancer is caused by physical agents, chemicals or viruses, the mechanism is the same the activation of oncogenes that are either native to the cell or introduced in viral genomes.

Oncogene = Gene responsible for a cell becoming cancerous.

· Discovered during the study of tumors induced by specific viruses.

· Harold Varmus and Micheal Bishop won a Nobel Prize for their discovery of oncogenes in RNA viruses (retroviruses) that cause uncontrolled growth of infected cells in culture.

Researchers later discovered that some animal genomes, including human, contain genes that

closely resemble viral oncogenes. These proto-oncognes normally regulate growth, division and adhesion in cells.

Proto-oncogenes = Gene that normally codes for regulatory proteins controlling cell growth, division and adhesion, and that can be transformed by mutation into an oncogene. Four types of mutations can convert proto-oncogenes to oncogenes:

1. Gene amplification. Sometimes more copies of oncogenes are present in a cell than is normal.

2. Chromosome translocation. Malignant cells frequently contain chromosomes that have broken and rejoined, placing pieces of different chromosomes side-by-side and possibly separating the oncogene form its normal control regions. In its new position, an oncogene may be next to active promoters or other control sequences that enhance transcription.

3. Gene transposition. Abnormal expression of an oncogene may occur if the oncogene or its regulatory gene are transposed to another locus. For example, a transposon may move:

· An oncogene to a new locus that has a highly active promoter.

· A promoter or other regulatory sequence to the upstream end of the oncogene.

4. Point mutation. A slight change in the nucleotide sequence might produce a growth stimulating protein that is more active or more resistant to degradation than the normal protein.

In addition to mutations affecting growth-stimulating proteins, changes in tumor supressor genes coding for proteins that normally inhibit growth can also promote cancer. For example,

· Children who are homozygous for the retinoblastoma allele develop malignant eye tumors.

· The mutated genes do not produce normal tumor-supressor protein.

· When a normal allele coding for tumor supressor is inserted into cells with mutated genes, abnormal growth of cancerous cells in culture is stopped.

· Mutant tumor-supressor gees are associated with inherited forms of colorectal cancer, Wilm’s tumor and breast cancer.

More than one somatic mutation is probably needed to transform normal cells into cancerous cells. One of the best understood examples is colorectal cancer.

· Development of metastasizing colorectal cancer is gradual, and the first sign is unusually rapid cell division of apparently normal cells in the colon lining: a benign tumor (polyp) appears, and eventually a malignant tumor may develop.

· During this process, mutation in oncogenes and tumor-supressor genes gradually accumulate. After a number of genes have changed, the tumor becomes mailgnant.

· In most cases studied, four changes occur at the DNA level: the activation of an oncogene (ras) and the inactivation of three tumor-supressor genes on chromosomes 5, 18, and 17.

Breast cancer, the second most common type cancer in women, is associated with somatic mutations of tumor-supressor genes.

· Inherited breast cancer accounts for 5-10% of all breast cancer cases.

· More than half of these are associated with mutations of a tumor-suppressor gene called BRCA1 ( for Breast Cancer) on chromosome #17.

· Another locus, BRCA2, on chromosome #13, accounts for most of the remaining breast cancer cases linked to family history.

· In breast cancer patients with no family history for the disease, new somatic mutations BRCA1 and BRCA2 are usually present.

· The study of such genes associated with breast cancer may lead to early diagnosis and treatment.

How are oncogenes involved in virus-induced cancers, which probably account for 15% of human cancer worldwide?

· Viruses might add oncogenes to cells or disrupt DNA, affecting proto-oncogenes or a tumor-supressor gene.

· However, viral-induced DNA changes are probably not all that is required to cause cancer, additional DNA changes needed may be triggered by environmental agents.