You are on page 1of 5

AP BIOLOGY REVIEW: Genetics Heredity: Eukaryotic Chromosomes How is genetic information organized in the eukaryotic chromosome?

Eukaryotic chromosomes contain an enormous amount of DNA relative to their condensed length. Each chromosome contains a single linear DNA double helix. Proteins called histones are responsible for the first level of DNA packing in chromosomes. Because they have a high proportion of positively charged amino acids, they bind tightly to negatively charged DNA. Linker DNA and histone molecules form beads on the extended chromatin fiber. A nucleosome consists of DNA wound around a protein core composed of two molecules each of four types of histone (H2A, H2B, H3, H4). The interaction between histone tails and linker DNA cause the chromatin fiber to coil or fold into the 30 nm fiber. This fiber forms looped domains, forming a 300 nm fiber. The chromatin folds further, resulting in the maximally compact chromosome seen in metaphase. How does this organization contribute to both continuity of and variability in the genetic information? According to the chromosome theory of inheritance, each Mendelian gene has a specific locus on a chromosome, and the chromosomes undergo independent assortment and segregation. Each chromosome has hundreds or thousands of genes. Genes located on the same chromosome that tend to be inherited together are linked genes. However, crossing over can occasionally break the physical connection between two genes on the same chromosome. During crossing over, the end portions of two nonsister chromatids trade places and bring a new combination of alleles. Molecular Genetics: Gene Regulation What are some mechanisms by which gene expression is regulated in prokaryotes and eukaryotes? Operon: A set of genes and the switches that control the expression of those genes. Ex) The Lac operons is switched off until it is induced to turn on. In order for transcription to occur, RNA polymerase must bind at the promotor. The relationship between RNA polymerase and the repressor is an example of noncompetitive inhibition. If allactose is present in the environment, it acts as an inducer, binding with the repressor so that it cannot bind to the operator. Another operons is the tryptophan operon, which is continuously switched on. When an inactive repressor combines with a tryptophan, it changes its conformation and binds to the operator, blocking RNA polymerase and transcription. In histone acetylation, acetyl groups (-COCH3) are attached to positively charged lysines in histone tails; deacetylation is the removal. When histone tails of a nucleosome are acetylated, the positive charges are neutralized and no longer bind to neighboring nucleosomes.

Transcription proteins have easier access to genes in an acetylated region. Histone acetylation enzymes may promote the initiation of transcription also by binding to recruiting components of transcription machinery. DNA methylation: The addition of methyl groups to certain bases in DNA after DNA is synthesized. Removal of extra methyl groups can turn on certain genes and researchers have discovered that certain proteins that bind to methylated DNA recruit histone deactylation enzymes. Thus, methylation and histone deacetylation can repress transcription. In some species, DNA methylation seems to be essential for long-term inactivation of genes that occurs during normal cell differentiation. Once methylated, enzymes correctly methylate the daughter strand after each round of replication. Epigenetic imprinting: inheritance of traits transmitted by mechanisms not directly involving the nucleotide sequence. Associated with most eukaryotic genes are multiple control elements, segments of noncoding DNA that help regulate transcription by binding certain proteins. These control elements and the proteins they bind are critical to the precise regulation of gene expression seen in different cell types. To initiate transcription, eukaryotic RNA polymerase requires the assistance of proteins called transcription factors. Only a few general transcription factors independently bind a DNA sequence, such as the TATA box. The others primarily bind to proteins, including each other and RNA polymerase II. Only when the complete initiation complex has assembled can the polymerase begin to move along the DNA template strand, producing a complementary strand of RNA. The interaction of general transcription factors and RNA polymerase II with a promoter usually leads to only a low rate of initiation and production of few RNA transcripts. Each eukaryotic gene has a promoter, a DNA sequence where RNA polymerase binds and starts transcription, proceeding downstream. Some control elements are named proximal control elements because they are located close to the promoter. The more distant distal control elements, groups of which are called enhancers, may be thousands of nucleotides upstream or downstream of a gene or even within an intron. An activator is a protein that binds to an enhancer and stimulates transcription of a gene. Protein-mediated bending of the DNA is thought to bring the bound activators in contact with a group of mediator proteins, which in turn interact with proteins at the promoter. These multiple protein-protein interactions help assemble and position the initiation complex on the promoter. Some specific transcription factors function as repressors to inhibit expression of a particular gene. Certain repressors block the binding of activators either to their control elements or to components of the transcription machinery. Other repressors bind directly to their own control elements in an enhancer and act to turn off transcription even in the presence of activators. Repressors and activators

can act indirectly, influencing chromatin structure. Some activators recruit proteins that acetylate histones near the promoters of specific gene, thus promoting transcription. Some repressors recruit proteins that deactylate histones, leading to reduced transcription- this is called silencing. Unlike genes in prokaryotic operons, each eukaryotic gene in these clusters has its own promoter and is individually transcribed. The coordinate regulation of clustered genes is thought to involve changes in the chromatin structure that make the entire group of genes either available or unavailable for transcription. More commonly co-expressed eukaryotic genes, such as genes coding for the enzymes of a metabolic pathway, are found scattered over different chromosomes. In these cases, coordinate gene expression seems to depend on the association of a specific control element or combination of elements with every gene of a dispersed group. Copies of the activators that recognize these control elements bind to them, promoting simultaneous transcription of the genes, no matter where they are in the genome. Coordinate control of dispersed genes in a eukaryotic cell often occurs as a response to external chemicals signals, such as a steroid hormone. Every gene whose transcription is stimulated by a particular steroid hormone, regardless of its chromosomal location, has a control element recognized by that hormone receptor complex. Other signaling molecules may initiate a signal transduction pathway that activates particular transcription activators or repressors. In Alternative RNA splicing different mRNA molecules are produced from the same primary transcript, depending on which RNA segments are treated as exons and introns. Regulatory proteins specific to a cell type control intron and exon choices by binding to regulatory sequences within the primary transcript. Initiation of translation of selected mRNAs can be blocked by regulatory proteins that bind to specific sequences or structures within the untranslated region at 5' end of mRNA, preventing the attachment of ribosomes. Another mechanism for blocking translation is seen in a variety of mRNAs present in egg cells: stored mRNAs lack poly-A tails big enough to initiate translation. Also, translation of all mRNAs in a cell may be regulated simultaneously. This involves activation or inactivation of one or more of the protein factors required to initiate translation. Some plants and algae store mRNAs during periods of darkness and light triggers the reactivation of translational apparatus. After translation, eukaryotic polypeptides must be processed to yield functional protein molecules. Also, many proteins undergo chemical modifications that make them functional. Regulatory proteins activated/inactivated by reversible addition of phosphate groups, and proteins destined for surface of animal cell acquire sugars. Some proteins must be transported to target destinations in cell to function.

Nucleic acid technology and applications

What are some current recombination technologies? Natural: viral transduction, bacterial transformation, conjugation, transposons DNA cloning: The plasmid is first isolated from a bacterial cell, and then the foreign DNA is inserted into it. The resulting plasmid is now a recombinant DNA molecule. The plasmid is returned to a bacterial cell (vector), producing a recombinant bacterium. Because the dividing bacteria replicate the recombinant plasmid and pass it on to their descendants, the foreign gene is cloned at the same time. Restriction Enzymes: enzymes that cut DNA molecules at a limited number of specific locations. Each restriction enzyme is very specific, recognizing a particular short DNA sequence, or restriction site, and cutting both DNA strands at specific points within this restriction site. The most useful restriction enzymes cleave the sugar-phosphate backbones in both DNA strands in a staggered way, resulting in double-stranded restriction fragments that have at least one single-stranded end, called a sticky end. These short extensions can form hydrogen-bonded base pairs with complementary sticky ends on any other DNA molecules cut with the same enzyme. The associations between fragments can be made permanent by the enzyme DNA ligase, which catalyzes the formation of covalent bonds that close up the sugar-phosphate backbones. Gel Electrophoresis: separates large molecules of DNA on the basis of their rate of movement through an agarose gel in an electric field. It is also used to separate proteins and amino acids. If DNA is going to be run through the gel, it must be cut up by restriction enzymes into small enough pieces. DNA can be sequenced based on the separation in the gel. A DNA probe can be used to identify the location of a specific sequence. DNA probe: radioactively labeled sing strand of nucleic acid molecule used to tag a specific sequence in a DNA sample. It bonds to the complementary sequence, and the radioactivity allows scientists to find it. (used to identify genetic defects like Huntingtons) Polymerase Chain Reaction: cell free, automated technique by which a piece of DNA can be rapidly copied. The DNA piece is put in a test tube with Taq polymerase along with nucleotides and primers. Once amplified, the copies can be compared with other samples. Restriction Fragment Length Polymorphisms (RFLPs): Each persons RFLPs are unique and inherited in Mendelian fashion. They are used in paternity suits or murder cases. cDNA: Use of reverse transcriptase to make DNA transcripts of RNA when scientists try to clone a human gene into a bacterium. What are some practical applications of nucleic acid technology? To produce a protein product (like insulin) for pharmaceutical use Gene therapy- replace a nonfunctioning gene Prepare gene copies for analysis Engineer bacteria to clean up environment

Diagnosing, treating, and even preventing diseases Vaccine production Crime investigation Genetic engineering has great potential to improve nutritional value of crop plants. ex) "golden rice" preventing vitamin A deficiency. Scientists have produced yellow rice plants containing beta-carotene, which the body uses to make vitamin A. Developing plants that make human proteins for medical use and viral proteins for use in vaccines. Large amounts of proteins may be produced more economically by plants than by cultured cells.

What legal and ethical problems may arise from these applications? Safety: o Much of the milk available comes from cows that have been given a genetically engineered bovine growth hormone to increase the quantity of milk produced. Privacy: o DNA probes are being coupled with the technology of the semiconductor industry to produce DNA chips that hold information about someones genetic makeup Health insurance companies or employers could show discrimination against people who are prone to genetic diseases Eugenics

You might also like