Topic 6 Flashcards
What is the central dogma of biological change of DNA to protein?
Information stored in DNA is unidirectionally transferred to RNA molecules during transcription and to proteins during translation
Slides 3-7
Slide 17 as we understand it today
Can RNA self replicate?
Can it be retrotranscribed to DNA?
RNA can replicate itself
RNA dependant RNA polymerase activity in RNA viruses
RNA can be retrotranscribed to DNA
Retrotranscriptase activity in retroviruses
Slides 9-12
Who is the original self replicator?
RNA
can store info (primary function of DNA)
Can transduce DNA messages into protein (primary function of RNA)
Can read info and catalyze reactions as ribozimes (primary function of proteins)
Slide 18
Overview the processes of transcription and translation in prokaryotes and eukaryotes on slides 20-21
Ok
How is the information in the DNA mechanistically transmitted to proteins in cells?
Transcription/RNA processing/translation
- Gene expression (RNA synthesis)
- mRNA processing (5’ capping, polymerization of poly A tail, intron splicing)
- mRNA transport
- mRNA translation
Slide 24
How is RNA synthesized?
RNA is synthesized in the nucleus, transported to the cytoplasm
Pulse chase labeling
Labeled RNA is exclusively in the nucleus
Later the labeled RNA is found in the cytoplasm
RNA synthesized in the nucleus and then transported to the cytoplasm
Slide 25
What are the 5 different RNA molecules involved in translation?
Messenger RNA (mRNA)- intermediates that carry generic info from DNA to ribosomes Small nuclear RNA (snRNA)- structural components of spliceosomes Transfer RNA (tRNA)- adaptors between amino acids and the codons in mRNA Ribosomal RNA (rRNA)- structural and catalytic components of ribosomes Micro RNA (miRNA)- short single stranded RNAs that block expression of complementary mRNAs
Slide 26
What are the features of RNA synthesis?
Similar to DNA synthesis except the precursors are ribonucleoside triphosphates (not deoxy), only one strand of DNA is used as template, RNA chains can be initiated de novo, uracyl instead of thymine
The RNA molecule will be complementary to the DNA template (antisense) strand and identical to the DNA nontemplate (sense) strand (except that uridine replaces thymidine)
RNA synthesis is catalyzes by RNA polymerases and ALWAYS proceeds in the 5’->3’ direction
Slide 27-34
What are the stages of transcription in prokaryotes?
- RNA chain initiation
- RNA chain elongation
- RNA chain termination
Slide 35
What is the first stage of transcription?
RNA polymerase binding to promoter
Initiation
After binding of RNA polymerase holoenzyme to promoter, localized unwind of 2 DNA strands by RNA polymerase to provide single stranded template
Formation of phosphodiester bonds between the first few ribonucleotides in the nascent RNA chain
Slides 36-39
What is the second stage of transcription?
Elongation
Rewinds the 2 DNA strands after done replicating new RNA strand
Slide 40
What is the 3rd stage of transcription?
Termination
Intrinsic mechanism
RNA being released
Rho dependant terminators- require a protein factor (ρ)
Ribosomes occupy transcript, cannot bind, stop codon in mRNA triggers release of attached ribosomes opening room for rho to bind, rho migrates on RNA and reaches polymerase where it rewinds the DNA closing the bubble
Rho independent terminators- do not require ρ
Slides 41-49
What are the 3 post transcriptional modifications (editing) of mRNAs?
Intron splicing
5’ capping
3’ poly-adenylation
What are the differences between prokaryotic and eukaryotic transcription?
One RNA polymerase vs 3 RNA polymerases
Sigma factors vs multi subunit general transcription factors
Co transcriptional translation vs spatially separated transcription and translation
Simple transcript vs processed (cap, introns/Exons, poly A tail)
DNA (almost naked) vs chromatin
Slide 51
What are the challenges in eukaryotic transmission?
Harder to locate the promoter, genome is bigger, genes are more spaced out
Transcription and translation are decoupled- they take place in different cellular components (nucleolus and cytoplasm)
Eukaryotic DNA is wrapped up around proteins- histones that need to be removed/moved for transcription to occur
Eukaryotic transcription is more complex- holoenyme has more subunits, transcriptional factors are required to recruit the polymerase and there are several types of RNA polymerases
Slide 52