RNA PROCESSING

Gyan Sundar Sahukhal
©1998 Timothy G. Standish

©1998 Timothy G. Standish

Standish .©1998 Timothy G.

 The most extensive processing of primary transcripts occurs in eukaryotic mRNAs and in tRNAs of both bacteria and eukaryotes.RNA Processing Many of the RNA molecules in bacteria and virtually all RNA molecules in eukaryotes are processed to some degree after synthesis. Standish .  A newly synthesized RNA molecule is called a primary transcript.  Some of the most interesting molecular events in RNA metabolism occur during this postsynthetic processing.  ©1998 Timothy G.

the introns are removed from the primary transcript and the exons are joined to form a continuous continuous sequence that specifies a functional polypeptide Eukaryotic mRNAs are also modified at each end. A modified residue called a 5 cap is added at the 5 end.” The primary transcript is processed. The coding segments are called exons In a process called splicing. and delivered to the ribosome for translation. Standish . ©1998 Timothy G. transported to the cytoplasm. The 3 end is cleaved.       Noncoding tracts that break up the coding region of the transcript are called introns. and 80 to 250 A residues are added to create a poly(A) “tail.

  Formation of the primary transcript and its processing during maturation of mRNA in a eukaryotic cell. A noncoding sequence following the last exon is shown in orange. ©1998 Timothy G. The 5 cap (red) is added before synthesis of the primary transcript is complete. Standish . Splicing can occur either before or after the cleavage and polyadenylation steps.

methylguanosine linked to the 5-terminal residue of the mRNA through an unusual 5.Eukaryotic mRNAs Are Capped at the 5 End Most eukaryotic mRNAs have a 5 cap.  The cap also binds to a specific cap binding complex of proteins and participates in binding of the mRNA to the ribosome to initiate translation  ©1998 Timothy G. Standish . a residue of 7.5triphosphate linkage  The 5 cap helps protect mRNA from ribonucleases.

     The 5 cap is formed by condensation of a molecule of GTP with the triphosphate at the 5 end of the transcript. after the first 20 to 30 nucleotides of the transcript have been added. All these reactions occur very early in transcription. and additional methyl groups are often added at the 2 hydroxyls of the first and second nucleotides adjacent to the cap. and through them the 5 end of the transcript itself. The capped 5 end is then released from the capping enzymes and bound by the cap-binding complex ©1998 Timothy G. Standish . are associated with the RNA polymerase II CTD until the cap is synthesized. All three of the capping enzymes. The guanine is subsequently methylated at N-7.

©1998 Timothy G. Standish .

but these tails stimulate decay of mRNA rather than protecting it from degradation. most eukaryotic mRNAs have a string of 80 to 250 A residues.  The poly(A) tail and its associated proteins probably help protect mRNA from enzymatic destruction.  This tail serves as a binding site for one or more specific proteins.  Many prokaryotic mRNAs also acquire poly(A) tails.Addition of poly A tail At 3’ end.  ©1998 Timothy G. making up the poly(A) tail. Standish .

2 The RNA is cleaved by the endonuclease at a point 10 to 30 nucleotides 3 to (downstream of) the sequence AAUAAA. and several other multisubunit proteins involved in sequence recognition. beginning at the cleavage site. Standish . ©1998 Timothy G. a polyadenylate polymerase.   1 The cleavage signal sequence is bound by an enzyme complex that includes an endonuclease. stimulation of cleavage. 3 The polyadenylate polymerase synthesizes a poly(A) tail 80 to 250 nucleotides long. and regulation of the length of the poly(A) tail.

©1998 Timothy G. Standish .

Standish .©1998 Timothy G.

©1998 Timothy G. Standish .

©1998 Timothy G. Standish .

©1998 Timothy G. Standish .

Standish .©1998 Timothy G.

and chloroplast genes coding for rRNAs. Group I and group II introns are also found among the rarer examples of introns in bacteria. The first two. differ in the details of their splicing mechanisms but share one surprising characteristic: they are self-splicing—no protein enzymes are involved. the group I and group II introns. ©1998 Timothy G. Standish . and plants. Group II introns are generally found in the primary transcripts of mitochondrial or chloroplast mRNAs in fungi. Group I introns are found in some nuclear. mRNAs. algae. mitochondrial.RNA Catalyzes the Splicing of Introns      There are four classes of introns. and tRNAs.

The splicing mechanisms in both groups involve two transesterification reaction steps  A ribose 2. Standish .  These reactions are very similar to the DNA breaking and rejoining reactions promoted by topoisomerases  ©1998 Timothy G. a new phosphodiester bond is formed at the expense of the old. maintaining the balance of energy.or 3-hydroxyl group makes a nucleophilic attack on a phosphorus and. in each step.

©1998 Timothy G. Standish .

Standish .Splicing mechanism of Group I intron ©1998 Timothy G.

Standish .Splicing mechanism of Group II intron ©1998 Timothy G.

Standish .©1998 Timothy G.

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