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Overexpression of essential splicing factor ASF/SF2 blocks the temporalshift in adenovirus pre-mRNA splicing and reduces virus progeny formation
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- Molin, Magnus (author)
- Uppsala universitet,Institutionen för medicinsk biokemi och mikrobiologi
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- Akusjärvi, Göran (author)
- Uppsala universitet,Institutionen för medicinsk biokemi och mikrobiologi
- (creator_code:org_t)
- 2000
- 2000
- English.
- In: Journal of Virology. - 0022-538X .- 1098-5514. ; 74:19, s. 9002-9009
- Journal article (peer-reviewed)
Abstract
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- Expression of cytoplasmic mRNA from most adenovirus transcription units is subjected to a temporal regulation at the level of alternative pre-mRNA splicing. The general tendency is that splice site selection changes from proximal to distal late after infection. Interestingly, ASF/SF2, which is a prototypical member of the SR family of splicing factors, has the opposite effect on splice site selection, inducing an increase in proximal splice site usage. We have previously shown that SR proteins late during an adenovirus infection become partially inactivated as splicing regulatory proteins. A prediction from these results is that overexpression of an SR protein, such as ASF/SF2, during virus growth will interfere with virus replication by disturbing the balance of functional and nonfunctional ASF/SF2 in the infected cell. To test this hypothesis, we reconstructed a recombinant adenovirus expressing ASF/SF2 under the transcriptional control of a regulated promoter. The results show that, as predicted, induction of ASF/SF2 during lytic virus growth prevents the early to late shift in mRNA expression from both early (E1A and E1B) and late (L1) transcription units. Furthermore, ASF/SF2 overexpression blocks viral DNA replication and reduces selectively cytoplasmic accumulation of major late mRNA, resulting in a lower virus yield. Collectively, our results provide additional support for the hypothesis that viral control of SR protein function is important for the proper expression of viral proteins during lytic virus growth.
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