Control of Eukaryotic Genes. AP Biology

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1 Control of Eukaryotic Genes

2 The BIG Questions How are genes turned on & off in eukaryotes? How do cells with the same genes differentiate to perform completely different, specialized functions?

3 Evolution of gene regulation Prokaryotes u single-celled u evolved to grow & divide rapidly u must respond quickly to changes in external environment exploit transient resources Gene regulation u turn genes on & off rapidly flexibility & reversibility u adjust levels of enzymes for synthesis & digestion

4 Evolution of gene regulation Eukaryotes u multicellular u evolved to maintain constant internal conditions while facing changing external conditions homeostasis u regulate body as a whole growth & development w long term processes specialization w turn on & off large number of genes serve the needs of the body as a whole rather than the needs of individual cells

5 Points of control The control of gene expression can occur at any step in the pathway from gene to functional protein 1. unpacking DNA 2. transcription 3. mrna processing 4. mrna transport out of nucleus through cytoplasm w protection from degradation 5. translation 6. protein processing 7. protein degradation

6 1. DNA packing How do you fit all that DNA into nucleus? u DNA coiling & folding double helix nucleosomes chromatin fiber looped domains chromosome from DNA double helix to condensed chromosome

7 Nucleosomes Beads on a string u 1 st level of DNA packing u histone proteins 8 protein molecules many positively charged amino acids w arginine & lysine bind tightly to negatively charged DNA 8 histone molecules

8 DNA packing as gene control Degree of packing of DNA regulates transcription u tightly wrapped around histones no transcription genes turned off heterochromatin darker DNA (H) = tightly packed euchromatin lighter DNA (E) = loosely packed H E

9 DNA methylation Methylation of DNA blocks transcription factors u no transcription genes turned off u attachment of methyl groups ( CH 3 ) to cytosine C = cytosine u nearly permanent inactivation of genes ex. inactivated mammalian X chromosome = Barr body

10 Histone acetylation Acetylation of histones unwinds DNA u loosely wrapped around histones enables transcription genes turned on u attachment of acetyl groups ( COCH 3 ) to histones conformational change in histone proteins transcription factors have easier access to genes

11 Epigenetics Epigenetics refers to processes that influence gene expression or function without changing the underlying DNA sequence. 1. Acetylation 2. Methylation Reading Homework on Epigenenomics Show the Epigentics PDF 11

12 Acetylation Acetylation of lysine found on the histone decreases the affinity of histones for DNA and other histones, thereby making DNA more accessible for transcription. 12

13 Methylation A methyl group can be added to the nitrogenous bases of cytosine that are followed by guanine. Many human genes have upstream CG-rich regions called CpG islands. Methylation of a gene's CpG island represses gene expression. Different cells have different methylation patterns, which contributes to the differences in gene expression in different cell types. 13

14 2. Transcription initiation Control regions on DNA u promoter nearby control sequence on DNA binding of RNA polymerase & transcription factors base rate of transcription u enhancers distant control sequences on DNA binding of activator proteins enhanced rate (high level) of transcription

15 Model for Enhancer action Enhancer DNA sequences u distant control sequences Activator proteins u bind to enhancer sequence & stimulates transcription Silencer proteins u bind to enhancer sequence & block gene transcription

16 Transcription complex Activator Proteins regulatory proteins bind to DNA at distant enhancer sites increase the rate of transcription Enhancer Sites regulatory sites on DNA distant from gene Enhancer Activator Activator Coactivator A B F E TFIID H T A T A Activator RNA polymerase II Coding region Core promoter and initiation complex Initiation Complex at Promoter Site binding site of RNA polymerase

17 3. Post-transcriptional control Alternative RNA splicing u variable processing of exons creates a family of proteins

18 4. Regulation of mrna degradation Life span of mrna determines amount of protein synthesis u mrna can last from hours to weeks

19 RNA interference Small interfering RNAs (sirna) u short segments of RNA (21-28 bases) bind to mrna create sections of double-stranded mrna death tag for mrna w triggers degradation of mrna u cause gene silencing post-transcriptional control, but still effectively turns off the gene = no protein produced sirna NEW!

20 sirna: RNA Interference (RNAi) Small single-standed RNA molecules called micrornas (mirnas) form complementary sequences by folding back on themselves. These hairpin structures are held together by H bonds. Enzymes called DICER trim these structures into small 20 nucleotide size pieces. One strand is degraded. The other which is the mirna binds to some proteins forming the mirna-protein complex called RISC or RNA induced silencing complex. These complexes can bind to any mrna that has a complementary sequence and can degrade that mrna, blocking translation. sirna or small interfering RNA are similar to mirna and they can both degrade mrnas by the same mechanism. sirnas and mirnas differ in their precursor molecules that generate them. These RNAi pathways can destroy viruses that have double-stranded RNA thus protecting the cell against viral infections.

21 Action of sirna dicer enzyme mirna or microrna mrna for translation Hot Hot new topic in biology sirna breakdown enzyme (RISC) double-stranded mirna + sirna mrna degraded functionally turns gene off

22 rnai/animation/index.html

23 5. Control of translation Block initiation of translation stage u regulatory proteins attach to specific sequences within the 5 UTR that is present at the 5 end of the mrna. This prevents attachment of ribosomal subunits & initiator trna and block translation of mrna to protein

24 6-7. Protein processing & degradation Protein processing u folding, cleaving, adding sugar groups, targeting for transport Protein degradation u ubiquitin tagging u proteasome degradation

25 Ubiquitin 1980s 2004 Death tag u mark unwanted proteins with a label u 76 amino acid polypeptide, ubiquitin u labeled proteins are broken down rapidly in "waste disposers" proteasomes Aaron Ciechanover Israel Avram Hershko Israel Irwin Rose UC Riverside

26 Proteasome Protein-degrading machine u cell s waste disposer u breaks down any proteins into 7-9 amino acid fragments cellular recycling

27 Proteasome-Degradation-of-Proteins

28 6 7 fate of protein after translation Gene Regulation 1 & 2. transcription - DNA packing - transcription factors 1 2 initiation of transcription 5 initiation of translation 3 & 4. post-transcription - mrna processing 4 - splicing - 5 cap & poly-a tail mrna destruction - breakdown by sirna 5. translation - block start of translation 6 & 7. post-translation - protein processing - protein degradation 3 mrna splicing 4 mrna protection

29 Turn your Question Genes on!

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