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Lecture 9 Transcription

The document discusses the process of transcription in molecular biology, detailing how RNA is synthesized from DNA through initiation, elongation, and termination phases. It highlights the roles of various transcription factors, the significance of capping and splicing in RNA processing, and the distinction between exons and introns. Additionally, it explains the function of the spliceosome and the concept of alternative splicing in generating diverse mRNA variants.

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3 views

Lecture 9 Transcription

The document discusses the process of transcription in molecular biology, detailing how RNA is synthesized from DNA through initiation, elongation, and termination phases. It highlights the roles of various transcription factors, the significance of capping and splicing in RNA processing, and the distinction between exons and introns. Additionally, it explains the function of the spliceosome and the concept of alternative splicing in generating diverse mRNA variants.

Uploaded by

césar mpassi
Copyright
© © All Rights Reserved
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
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LECTURE 9

Заголовок
Цифровая 3D-медицина
TRANSCRIPTION: RNA SYNTHESIS
Результаты в области компьютерной графики и геометрического моделирования
Подзаголовок презентации

Dr. Svetlana V. Trofimova


2021
The simplified scheme of the central dogma

The central dogma of molecular biology is DNA codes for RNA, which codes for proteins.
A gene is a section of DNA that codes for a particular protein and determines a certain trait. A gene is the basic unit of heredity.
Transcription is a process in which ribonucleic acid (RNA) is synthesized from DNA.
The common features of transcription and replication are:
their fundamental chemical mechanism
• polarity (direction of synthesis)
• use of a template.
• like replication, transcription has initiation, elongation, and termination phases.

Transcription differs from replication in that it does not require a primer and, generally, involves only limited segments of a
DNA molecule. Additionally, within transcribed segments only one DNA strand serves as a template for a particular RNA
molecule.

DNA dependent RNA polymerase (RNA pol) is an enzyme that is responsible for copying a DNA sequence into an RNA
sequence during the transcription.
The RNA is synthesized from a single strand or template/sense strand of a DNA molecule.
The opposite DNA strand is called a coding strand, because its nucleotide sequence corresponds to the sequence of the RNA
transcript produced.
The section of DNA that is transcribed into an RNA molecule is called a transcription unit.
A transcription unit codes the sequence that is translated into protein.
Promoter sequences are DNA sequences that define where transcription of a gene by RNA polymerase begins.
Terminator - the nucleotide sequence of DNA on which gene transcription is completed.

Transcription occurs in three sequential stages: initiation, elongation, and termination.


Stages of Transcription: Initiation
Initiation occurs when RNA polymerase binds at specific DNA sequences upstream from the gene that will be transcribed
called promoters. The structure of eukaryotic promoters is generally complex, and they have several different sequence
motifs, which include the TATA-box, initiator (Inr-box), CAAT-box and GC-box.
Transcription factors (TFs) are the proteins that help determine which DNA sequences should be transcribed and
precisely when the transcription process should occur.

The first base on the DNA where transcription actually starts is labeled +1. Sequences that are upstream of the first base
of the transcript are marked with negative numbers. Sequences that are downstream of the first base of the transcript are
marked with positive numbers.
Stages of Transcription: Initiation
Summary of eukaryotic RNA polymerase II general transcription factors
TF Number of subunits Function
TFIIA 3 Stabilizes binding of TATA-binding protein (TBP) and TFIIB
TFIIB 1 Binds TBP, selects start site, recruits RNA pol II
TFIID 12 Interacts with regulatory factors
TBP 1 Subunit of TFIID, specifically recognizes the TATA box
TFIIE 2 Recruits TFIIH
TFIIF 2 Binds RNA pol II and TFIIB
TFIIH 9 Unwinds promoter DNA, phosphorylation of RNA pol II
RNA poll II 12 Catalyzes RNA synthesis

Simplified model of protein complex assembly on DNA is shown


Stages of Transcription: Elongation

The RNA pol moves stepwise along the DNA, unwinding the DNA helix at its active site. It adds nucleotides one by one
to the RNA chain at the polymerization site using an exposed DNA strand as a template. The incoming nucleotides are in
the form of ribonucleoside triphosphates (ATP, UTP, CTP, and GTP), and the energy stored in their phosphate-phosphate
bonds provides the driving force for the polymerization reaction
mRNA Capping

Capping is the first modification made to RNA and takes place co-
transcriptionally in the nucleus when the first 25-30 nucleotides are
incorporated into the nascent
transcript. Three enzymes, RNA triphosphatase, guanylyltransferase,
and methyltransferase are involved in the addition of the cap to the
mRNA. Capping of the pre-mRNA is the addition of 7-
methylguanosine (m7G) to the 5'-end. Capping the 5'-end of the RNA
transcript largely determines its further fate in the cell. The following
cap functions are known:
• regulation of transcription;
• participation in splicing;
• participation in the processing of the 3'-end of mRNA;
• regulation of RNA transport between the nucleus and the cytoplasm;
• protection of the transcript from degradation by exonucleases;
• translation stimulation.
Stages of Transcription: Termination and Polyadenylation

The stage of termination begins when RNA polymerase reaches the specific nucleotide sequence referred to
as termination site, the elongation factor being separated from and termination factor bound to RNA polymerase. It
facilitates separation of newly synthesized pre-RNA molecule and enzyme from DNA template.

Transcriptional termination of eukaryotic mRNA genes occurs when RNA pol II encounters specific termination
signals. The canonical polyadenylation signal (PAS) 3'-AAUAAA-5' (or less frequently AUUAAA) directs the
incorporation of the termination and polyadenylation signal.
RNA splicing

Exons are coding regions of genes.


Introns are non-coding regions of genes.
Splicing is the excision of copies of introns from pre-mRNA and stitching of copies of exons to form mRNA.
Pre-mRNA is the first transcript of a gene that encodes proteins (precursor mRNA).
RNA processing is a set of processes in eukaryotic cells that lead to the transformation of the primary transcript into
mature RNA.
Splicing is the process of removing certain nucleotide sequences from RNA molecules and joining the sequences that are
stored in the "mature" molecule during RNA processing.
The spliceosome is a nuclear structure consisting of RNA molecules and proteins and which removes non-coding
sequences from mRNA precursors.
Alternative splicing is a variant of messenger RNA splicing, in which several mature mRNAs are formed during gene
expression based on the same primary transcript.
RNA splicing
The spliceosome is assembled from small nuclear
ribonucleoproteins (snRNPs). snRNPs recognize conserved
sequences in splice sites, remove the introns, and past the
exons together. (A) Schematic diagram of the steps of
spliceosome assembly and intron removal. Introns are
removed by RNA processing in which the intron is looped
out and cut away from the exons by snRNPs, and the exons
are spliced together to produce the translatable mRNA (B).
When different coding regions of mRNA are spliced out,
different variations of matured mRNAs that encode different
proteins will eventually occur

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