Lecture RNA synthesis Flashcards
RNA is a polymer composed of alternating units of ribonucleotides connected through a
3-5 phospodiester bond
RNA is a polymer composed of alternating units of
Ribonucleosides
In contrast with DNA, ribonucleotides contain
hydroxyl groups on the 2’-carbonof the ribose sugar •
the base uracilin place of thymine
The “working copies” of DNA •Expresses the master plan contained in DNA
Significance of RNA
80% of the total RNA
Ribosomal RNA
smallest of the 3 major RNA species (excluding the small RNAs), make up 15%
Transfer RNA
2-5%, but is the most heterogenousin terms of size and base sequence
Messenger RNA
involved in mRNA splicing and gene regulation
Small RNA’s
Prokaryotic rRNA
23S, 16S, 5S
Eukaryotic rRNA
28S, 18S, 5.8S, 5S
measure of size based upon the molecular sedimentation rate during ultracentrifugation
Svedberg unit “S”
Similarities between DNA and RNA
Both have the general steps of initiation, elongation, and termination with 5’ to 3’ polarity (synthesized in a 5’ 3’ direction, antiparallelto the DNA template strand which is read in a 3’ 5’ direction) •Both have large, multicomponentinitiation complexes •Both adhere to Watson-Crick base-pairing rules
RNA
DNA
Ribose
Deoxyribose
complementary base pair for A in RNA
U instead of T (for DNA)
not involved in RNA synthesis
Primer
Only a portion of the genome is transcribed or copied
RNA
entire genome must be copied
DNA
always read in the 3’ to 5’ direction (RNA)
Template strand
The opposite strand is called the (RNA)
Coding strand
The enzyme responsible for the polymerization of ribonucleotidesinto a sequence complementary to the template strand of the gene
DNA dependent RNA polymerase
DNA DEPENDENT RNA POLYMERASE attach to the ______ on the ______
Promoter
Template strand
corresponds to the 5’ nucleotide of the mRNA
Starting point of transcription
The numbers increase as the sequence proceeds
Downstream
The nucleotide in the promoter adjacent to the transcription initiation site is designated -1, and these negative numbers increase as the sequence proceeds
Upstream
defined as that region of DNA that includes the signals for transcription: initiation, elongation, and termination
Transcription unit
A transcription unit includes a
Promoter
RNA coding region
Terminator
The RNA product, which is synthesized in the 5’ to 3’ direction, is the
Primary transcript
with its 4 subunits responsible for 5’ 3’ RNA polymerase activity
Core enzyme
Core enzyme cannot recognize the promoter on the DNA template
Lack specificity
contains the sigma subunit or sigma factor (that enables RNA polymerase to recognize promoter regions on the DNA
Holoenzyme
An auxiliary protein of RNA polymerase
Termination factor
Some regions of DNA that signal the termination of transcriptionare recognized by
RNA polymerase
Others are recognized by specific termination factors, an example of which is the
rho factor of ecoli
The primary transcripts generated by RNA polymerase II(one of three nuclear DNAdependent RNA polymerases in eukaryotes) are promptly capped by
7 methylguanosine triposphate
These caps are necessary for the subsequent processing of the primary transcript to mRNA, for the translation of the mRNA, and for protection of the mRNA against exonucleolyticattack
7 methylguanosine triposphate
Involves binding of the RNA polymerase holoenzymeto a promoter region
Initiation
Pribnowbox and -35 sequence •Highly conserved •Recognized by prokaryotic RNA polymerase sigma factors
Consensus nucleotide sequences
A stretch of 6 nucleotides (5’-TATAAT-3’) centered around 8 to 10 nucleotides to the left of the transcription start site that codes for the
Initial base of mRNA
A stretch of 6 nucleotides (5’-TATAAT-3’) centered around 8 to 10 nucleotides to the left of the transcription start site that codes for the initial base of mRNA
Pribnow box
A second consensus nucleotide sequence centeredaround 35 bases to the left of the transcription start site
-35 sequence
A mutationin either the Pribnowbox or -35 sequence can affect the transcription of the gene controlled by the
Mutant promoter
Once the promoter has been recognized by the holoenzyme, RNA polymerase (RNAP) begins to synthesize a transcript of the DNA sequence
Elongation
Elongation usually beginning with a
Purine
Unlike DNA polymerase, RNA polymerase does not require a
primer and has no endoor exonucleaseactivity (No repair capability
longation by addition of ribonucleotidesto the
3-OH end
3’-OH acts as a __________, attacking the -phosphate of the incoming ribonucleosidetriphosphate and releasing ___________
Nucleophile
Pyrophosphate
Alternatively, can recognize termination regions on the DNA template
tetramericRNA polymerase
may be required for the release of the RNA product
Rho factor protein