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RNA MODIFICATION
Most of eukaryotic mRNAs are spliced after transcription In eukaryotic genes, coding sequences, called exons, are usually interrupted by intervening sequences or introns Transcription produces the entire gene product Introns are later removed or excised Exons are connected together or spliced This phenomenon is termed RNA splicing It is a common genetic phenomenon in eukaryotes Occurs occasionally in bacteria as well

RNA MODIFICATION
Aside from splicing, RNA transcripts can be modified in several ways For example
Trimming of rRNA and tRNA transcripts 5 Capping and 3 polyA tailing of mRNA transcripts

Refer to Table 12.3

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Trimming
Many nonstructural genes are initially transcribed as a large RNA, and is enzymatically cleaved into smaller functional pieces

S: Svedberg unit used for sedimentation rate; here used for the molecule size of RNA

This processing occurs in the nucleolus

Figure 12.14

Functional RNAs that are key in ribosome structure

tRNAs are also made as large precursors, and will be cleaved at both the 5 and 3 ends to produce mature, functional tRNAs

Endonuclease

(Endonuclease) RNase P

(RNase D)

Found to contain both RNA and protein subunits However, RNA contains the catalytic ability Therefore, it is a ribozyme

Figure 12.15

mRNA Capping
In eukaryotes, the transcription of structural genes, produces a long transcript known as pre-mRNA Also as heterogeneous nuclear RNA (hnRNA) Pre-mRNA consisting of exons and introns is spliced (excising introns) to form mature mRNA Most mature mRNAs have a 7-methyl guanosine covalently attached at their 5 end This event is known as capping Capping occurs as the pre-mRNA is being synthesized by RNA pol II Usually when the transcript is only 20 to 25 bases long 12-64
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Removes one of the phosphates

Attaches GMP to the 5 end

Figure 12.19
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Attaches a methyl group to the guanine base

Figure 12.19
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mRNA Tailing
Most mature mRNAs have a string of adenine nucleotides at their 3 ends
This is termed the polyA tail

The polyA tail is not encoded in the gene sequence

It is added enzymatically after the gene is completely transcribed

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Figure 12.20

Consensus sequence in higher eukaryotes

Appears to be important in the stability of mRNA and the translation of the polypeptide

Length varies between species From a few dozen adenines to several hundred

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Splicing
Three different splicing mechanisms have been identified
Group I intron splicing Group II intron splicing Spliceosome

All three cases involve

Removal of the intron RNA Linkage of the exon RNA by a phosphodiester bond

Splicing among group I and II introns is termed self-splicing

Splicing does not require the aid of enzymes Instead the RNA itself functions as its own ribozyme
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Self splicing of group I and II introns

Self-splicing introns
Relatively uncommon Group I introns found in rRNA genes within the nucleus of Tetrahymena and other lower eukaryotes Group II introns found in structural, tRNA and rRNA genes within mitochondria DNA and in chloroplast DNA

Figure 12.18

Pre-mRNA splicing by spliceosome

The spliceosome is a large complex that splices pre-mRNA It is composed of several subunits known as snRNPs Each snRNP contains small nuclear RNA and a set of proteins The subunits of a spliceosome: Bind to an intron sequence and precisely recognize the intron-exon boundaries Catalyze the chemical reactions that remove introns and covalently link exons
Figure 12.16
Spliceosome

 Intron RNA is defined by particular sequences within the intron and at the intro-exon boundaries The consensus sequences for the splicing of mammalian pre-mRNA are shown in Figure 12.21
Sequences shown in bold are highly conserved Corresponds to the boxed adenine in Figure 12.22

Figure 12.21

Serve as recognition sites for the binding of the spliceosome

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Intron loops out and exons brought closer together

Figure 12.22
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Intron will be degraded and the snRNPs used again

Figure 12.22
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Intron Advantage?
Alternative splicing A pre-mRNA with multiple introns can be spliced in different ways to generate mature mRNAs with different combinations of exons
Exon I Intron I Exon II Intron II Exon III Intron III Exon IV

Splicing
Exon I Exon II Exon IV Exon I Exon II Exon III Exon IV

This variation in splicing can occur in different cell types or during different stages of development The biological advantage of alternative splicing is that two (or more) polypeptides can be derived from a single gene This allows an organism to carry fewer genes in its genome

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