Functional RNAs are produced through modification of pre-RNA produced
from transcription process except bacterial mRNAs which are as such used for
protein synthesis without any modification. These series of modification
involving removal of introns by splicing are known as processing of RNA.
of rRNA & tRNA:
The processing of both rRNA & tRNA in prokaryotes and
eukaryotes are similar. Eukaryotes – 4 species of rRNAs, three of which (28S,
18S & 5.8S) are derived by cleavage from single long precursor transcript(pre-rRNA).
Whereas the fourth(5S) is transcribed from a separate gene. There are 3 rRNAs
in prokaryotes(23S, 16S & 5S) which are also transcribed from a single
In prokaryotic cell initial cleavage yield separate precursor of 3
individual rRNAs which undergo secondary cleavage to produce functional forms. Whereas
in eukaryotic cell(within nucleolus) initial cleavage near 5′ side of 5.8S rRNA
results in 2 segments(18S & 28S + 5.8S). Further cleavage produce
functional forms with 5.8S having hydrogen-bonded to 28S. Apart from this there
will be also some addition of methyl groups to the bases & sugar moieties
of specific nucleotides and conversion of some uridines to pseudouridines.
Similarly from pre-tRNAs individual tRNA are synthesised in both
prokaryotes & eukaryotes. In prokaryotes, some tRNA s are included in
RNase P enzyme(ribozyme) involved in processing at 5′ end of pre-tRNAs. It
consists of RNA & protein molecules. Here RNA is responsible for catalytic
activity whereas proteins are required for maximal activity.
3′ end processing involves action of conventional protein RNase and
addition of a CCA terminus. As this CCA sequences are the site of protein
attachment, it is present in all tRNAs for protein synthesis. In addition approximately
10% of bases of specific nucleotides are modified. Introns are spliced using
endonuclease from pre-tRNAs.
Processing of mRNA in eukaryotes:
In contrast to prokaryotes, pre-mRNA synthesised in nucleus of
eukaryotes are modified before transported to cytoplasm. Processing involves
modification of both ends of initial transcript and removal of introns. The C-terminal
domain(CTD) of RNA polymerase II serve as binding site for enzyme complexes. Whereas
polymerases I and III lack a CTD, so their transcripts are not processed by
same enzyme complexes.
Processing at 5′ end – addition of 7-methylguanosine cap.
After transcription of first 20-30 nucleotides, enzymes for capping are
recruited to pCTD. Capping initiated by addition of a GTP in reverse
orientation to 5′ terminal nucleotide. Followed by addition of methyl group to
this G residue & to ribose moieties of one or two 5′ nucleotides. This 5′
cap helps stabilising & aligning on ribosomes duting protein synthesis.
At 3′ end – cleavage of primary transcript downstream to 10-30
nucleotides of highly conserved hexanucleotide(AAUAAA in mammalian cells) and
addition of a poly-A tail known as polyadenylation.
Thus G-U rich residue is degraded.