UNIVERSITY INSTITUTE OF BIOTECHNOLOGY

Master of science (zoology)

Subject name = B i o s t a t i s t i c s
Subject code= 2 3 S Z T - 6 1 5
Topic= transcription in prokaryotes and eukaryotes

presented by- sneha kumari

UID= 2 3 M S Z 1 0 0 4 6
Submitted to –Dr. Naveen kumar
Transcription in prokaryotes tr

T h i s Photo by Unknown author is licensed under C C B Y - S A - NC.

 3 s t e p s of transcription

• Transcription by E. coli R N A polymerase occurs in three phases;

• initiation, elongation and termination.
• Initiation involves binding of the enzyme to a promoter upstream of the gene.
• During elongation, the antisense D N A strand is used a s the template so that the
R N A made has the same base sequence a s the sense (coding) strand, except that U
replaces T.
• A termination signal is eventually encountered that halts synthesis and causes
release of the completed RNA.
 3 p h a s e s of gene transcription

• Three phases of Gene transcription by E. coli R N A polymerase takes place in three phases: transcription
initiation, elongation and termination.
• During initiation, R N A polymerase recognizes a specific site on the DNA, upstream from the gene that will be
transcribed, called a promoter site and then unwinds the D N A locally.
• During elongation the R N A polymerase u s e s the antisense () strand of D N A a s template and synthesizes a
complementary R N A molecule using ribonucleoside 5’ triphosphates as precursors. Th e R N A produced has the
same sequence as the non-template strand, called the s e n s e () strand (or coding strand) except that the R N A
contains U instead of T. At different locations on the bacterial chromosome, sometimes one strand is used as
template, the correct strand to be used as template is identified for the R N A polymerase by the presence of the
promoter site.
• Finally, the R N A polymerase encounters a termination signal and c e a s e s transcription, releasing the R N A
transcript and dissociating from the DNA.
 Initiation in prokaryotes
1 . R N A polymerase holoenzyme (containing
2 subunits) initiates transcription by binding to a 4 0 – 6 0
bp region that contains two conserved promoter
elements, the –10 sequence (Pribnow box) with the
consensus T A T A A T and the –35 sequence with the
consensus T T G A C A . The factor is essential for
initiation. T h i s Photo by Unknown author is licensed under C C BY.

2. No primer is required. Promoters vary up to 1000-fold

in their efficiency of initiation which depends on the
exactsequence of the key promoter elements a s well as
flanking sequences
Following initiation, the subunit dissociates from
R N A polymerase to leave the core enzyme (2 ) that
continues R N A synthesis in a 5 → 3 direction using
the four ribonucleoside 5 triphosphates as
precursors. Th e D N A double helix is unwound for
transcription, forming a transcription bubble, and is
then rewound after the transcription complex has
passed. A common termination signal is a hairpin
structure formed by a palindromic G C - rich region,
followed by an A T - rich sequence. Other signals are
also used which require the assistance of rho ()
protein for effective termination.
T h i s Photo by Unknown author is licensed under C C BY.
Prokaryotic promoter showing the –10 sequence and –35 sequence. By
convention, the first nucleotide of the template D N A that is transcribed into
R N A is denoted +1, the transcriptional start site
 In E. coli, all genes are transcribed by a single large R N A polymerase with the subunit
structure 2 . This complete enzyme, called the holoenzyme, is needed to initiate transcription
since the factor is essential for recognition of the promoter; it decreases the affinity of the
core enzyme for nonspecific D N A binding sites and increases its affinity for the promoter.

Within the promoter lie two 6 - bp sequences that are particularly important for promoter
function and which are therefore highly conserved between species. Using the convention of
calling the first nucleotide of a transcribed sequence a s 1, these two promoter elements lie
at positions –10 and –35, that is about 10 and 3 5 bp, respectively, upstream of where
transcription will begin (Fig. 1).
● T h e –10 sequence has the consensus T A T A A T . Because this element was discovered by
Pribnow, it is also known a s the Pribnow box. It is an important recognition site that interacts
with the σ factor of R N A polymerase.
● T h e –35 sequence has the consensus T T G A C A and is important in D N A unwinding during
transcriptional initiation. T h e actual sequence between the –10 sequence and the –35
sequence is not conserved (i.e. it varies from promoter to promoter) but the distance
between these two sites is extremely important for correct functioning of the promoter.
Promoters differ by up to 1000-fold in their efficiency of initiation of transcription so that
g enes with strong promoters are transcribed very frequently.
whereas genes with weak promoters are transcribed
far less often. The –10 and –35 sequences of strong
promoters correspond well with the consensus
sequences shown in Fig. 1 whereas weaker promoters
may have sequences that differ from these at one or
more nucleotides. The nature of the sequences around
the transcriptional start site can also influence the
efficiency of initiation. R N A polymerase does not need a
primer to begin transcription (cf. DN A polymerases,
T op ic s F 3 and F4); having bound to the promoter site,
the R N A polymerase begins transcription directly
Elongation in prokaryotes
 Elongation in prokaryotes

After transcription initiation, the σ factor is released from the transcriptional complex to leave the core
enzyme (α2ββ ω) which continues elongation of the RNA transcript. Thus the core enzyme contains
the catalytic site for polymerization, probably within the β subunit. The first nucleotide in the RNA
transcript is usually pppG or pppA. The RNA polymerase then synthesizes RNA in the 5 → 3
direction, using the four ribonucleoside5 triphosphates (ATP, CTP, GTP, UTP) as precursors. The 3 -
OH at the end of the growingRNA chain attacks the α phosphate group of the incoming
ribonucleoside 5 triphosphate to form a 3 5 phosphodiester bond (Fig. 2). The complex of RNA
polymerase, DNA template and new RNA transcript is called a ternary complex (i.e. three
components) and the region of unwound DNA that is undergoing transcription is called the
transcription bubble (Fig. 3). The RNA transcript forms a transient RNA–DNA hybrid helix with its
template strand but then peels away from the DNA as transcription proceeds. The DNA is unwound
ahead of the transcription bubble and after the transcription complex has passed, the DNA rewinds
 Termination in prokaryote

Transcription continues until a termination

sequence is reached. Th e most common
termination signal is a G C - rich region that is a
palindrome, followed by an A T - rich sequence.
Th e R N A made from the D N A palindrome is self
complementary and s o base-pairs internally to
form a hairpin structure rich in G C base pairs
followed by four or more U residues (Fig. 4).
However, not all termination sites have this
hairpin structure. Th ose that lack such a T h i s Photo by Unknown author is licensed under C C B Y - S A - NC.

structure require an additional protein, called

rho (ρ), to help recognize the termination site
and stop transcription
 R N A proces s ing

In prokaryotes, R N A transcribed from protein- coding genes

(messenger RNA, mRNA), requires little or no modification prior to
translation. In fact, many mRNA molecules begin to be
translated even before R N A synthesis has finished. However, ribosomal
R N A (rRNA) and transfer R N A (tRNA) are synthesized as precursor
molecules that do require post- transcriptional processing
THANK YOU