2011 BL 426 Study guide test 4 Ch 16, 19, 20, 21 other RNA processing, ribosome, tRNA, DNA replication, DNA repair telomere 1. The test will be worth 125 pts and will have a number of multiple choice questions, plus some matching probably, maybe gel or experiment to interpret, and some compare/contrast, or short essay type questions; maybe also some abbreviations to define. 1. Focus on what was presented in PowerPoints, using your text for additional clarifications and support. 2. Read learning outcomes for each chapter; read summary at end of each chapter for topics covered 3. Review the PowerPoint slides. 4. Focus on understanding important Figures – what is being shown; what type of experiment was done. 5. Consider suggested review and analytical questions at the end of chapters. 6. Make a list of important new terms, concepts, enzymes, DNA sites, RNA sites and what they do (many are listed below, but the list may not be comprehensive) 7. Complexity: Lots of big protein/RNA and protein/DNA complexes are involved 8. Consider different model systems (bacteria, yeast, mammals) and what they told us. Note that much of this material is about eukaryotes, although prokaryotic systems were critical for Xray crystallography of tRNA, of ribosome subunits. viruses (lambda, SV40), Tetrahymena telomeres, trypanosome RNA editing, Haloarcula, Thermus thermophilus, E. coli, C. elegans, Saccharomyces, 9. Some general comparisons of eukaryotes to prokaryotes – what is similar, what must be different? Consider RNA processing, DNA replication, repair, ribosomes, tRNA, mRNA; Mg++ was often a cofactor for cleavages DNA replication (circle, linear, telomeres) 10. Consider some of the types of experimental techniques and assays were used to obtain the understanding of the molecular mechanisms, many of which we have mentioned before. in vitro RNA synthesis; RNA and DNA gels; protein-RNA binding; telomerase assay Purification of proteins and protein complexes (affinity tags, ion exchange chromatography) EMSA; linker scanning mutagenesis; primer extension RNA synthesis; X-ray crystallography; mutated RNA sites; Mutant proteins, mutated DNA binding sites, Chromatin immunoprecipitation; how structure informs funciton Some new concepts: rRNA and tRNA processing; tRNA joined to amino acids trans-splicing vs. cis-splicing RNA editing RNAi Ribosome subunits structure and function; selectivity of aa-tRNA synthetase enzyme ,; telomerase; telomerase has guide RNA plus proteins Structures of telomeres and pecific proteins binding to end of telomeres DNA damage and repair of typical damage; better to repair before replication: Reversal of damage; Base excision repair, nucleotide excision repair, recombination repair Alkylating agents, thymine dimers, mismatch, double strand break; SOS repair Some DNA sequences and signals: oriC, ARS, Telomeres, centromeres, start of transcription Some jargon/ abbreviations: Pol II (CTD) 5’-UTR 3’-UTR Upstream downstream RFLP NER BER ORF XP Small subunit large subunit RNA start is +1 Some new, some review terms: processivity semi-conservative polysome leading strand lagging strand, polycistronic monocistronic rolling circle replication bidirectional replication sliding clamp Some protein complexes, what they do and what they contain: DNA polymerase III holoenzyme, primosome, primase RISC telomerase DNA polymerases Some proteins and enzymes: Ribozyme RNase T1 RNase III RNase A RNase T2 RNase E RNase P nuclease S1 DNase I Antibody RNaseH RdRP (RNA-dependent RNA polymerase) amino acyl tRNA synthetase TUTase (terminal uridyl transferase) Histone methyl transferase DNA pol I DNA pol III (and subcomplexes), Klenow polymerase DNA ligase Eukaryotic DNA pol: () main ones Photolyase DNA glycosylase AP endonuclease helicase Excinuclease (uvrABC) recA protease umuCD (DNA polV) TERT (telomerase RT) topoisomerase o6-meG methyltransferase T antigen of SV40 Dicer Ago peptidyl transferase Some RNA molecules: tRNA, rRNA, mRNA, 5S rRNA, snRNA, hnRNA, 16S 23S 18S 28S 5S 5.8S Guide RNA (editing) telomerase RNA siRNA miRNA guide RNA (RNAi)