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1 Center for Cancer and Immunology Research, Room 5720, Children's Research Institute, George Washington University School of Medicine and Health Sciences, 111 Michigan Avenue, NW, Washington, DC 20010, USA
2 Department of Infectious Diseases, Children's National Medical Center, George Washington University School of Medicine and Health Sciences, 111 Michigan Avenue, NW, Washington, DC 20010, USA
3 Department of Pediatrics, George Washington University School of Medicine and Health Sciences, 111 Michigan Avenue, NW, Washington, DC 20010, USA
Correspondence
Anamaris Colberg-Poley (at Children's Research Institute)
acolberg-poley{at}cnmcresearch.org
The human cytomegalovirus (HCMV) UL36–38 immediate early (IE) locus encodes proteins required for virus growth. The UL37 IE promoter drives production of differentially spliced and unspliced RNAs. To study their post-transcriptional processing, we generated target minigenes encoding each UL37 RNA splicing substrate. Target 1 RNA, spanning UL37 exon 1 (x1) donor and 2 (x2) acceptor as well as adjacent intronic sequences, but not the UL38 gene, accurately reproduced UL37 x1/x2 RNA splicing in transfected permissive cells. Surprisingly, deletion of distal intronic sequences nt -82 to -143 from the UL37x2 acceptor resulted in aberrant splicing to an upstream non-consensus exonic donor. Target 1 RNAs carry the UL37x1 polyadenylation (PA) signal and site as well as a downstream SV40 early PA signal. Both the UL37x1 and SV40 PA signals are used in wild-type target 1 RNAs but inhibited in UL37x1 PA signal mutants. Alternative RNA splicing of UL37 exons 2 to 3 or 3A as well as exons 3 to 4, observed in HCMV mature UL37 and UL36 spliced RNAs, is accurately reproduced with target minigene RNAs carrying the corresponding UL37 exonic and intronic sequences. Moreover, alternative splicing using two novel UL37 exon 3 consensus splice donors (di and dii) was found in target and in HCMV-infected cell RNA. These results demonstrate that: (i) target minigene RNAs accurately recapitulate the processing of UL37 IE RNAs in the HCMV-infected cell; (ii) precise UL37x1 donor selection is modulated by 3'-distal UL37 intronic sequences; and (iii) UL37 exon 3 contains multiple alternative consensus splice donors.
Present address: Center for Community-Based Health Strategies, Academy for Educational Development, 1825 Connecticut Ave, NW, Washington, DC 20009, USA.
Present address: Jordan University of Science and Technology, Irbid, Jordan.
This article has been cited by other articles:
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  M. S. Mavinakere, C. D. Williamson, V. S. Goldmacher, and A. M. Colberg-Poley Processing of Human Cytomegalovirus UL37 Mutant Glycoproteins in the Endoplasmic Reticulum Lumen prior to Mitochondrial Importation J. Virol., July 15, 2006; 80(14): 6771 - 6783. [Abstract] [Full Text] [PDF]
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R. Berro, K. Kehn, C. de la Fuente, A. Pumfery, R. Adair, J. Wade, A. M. Colberg-Poley, J. Hiscott, and F. Kashanchi Acetylated Tat Regulates Human Immunodeficiency Virus Type 1 Splicing through Its Interaction with the Splicing Regulator p32. J. Virol., April 1, 2006; 80(7): 3189 - 3204. [Abstract] [Full Text] [PDF]
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 R. Adair, G. W. Liebisch, Y. Su, and A. M. Colberg-Poley Alteration of cellular RNA splicing and polyadenylation machineries during productive human cytomegalovirus infection J. Gen. Virol., December 1, 2004; 85(12): 3541 - 3553. [Abstract] [Full Text] [PDF]
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M. S. Mavinakere and A. M. Colberg-Poley Internal cleavage of the human cytomegalovirus UL37 immediate-early glycoprotein and divergent trafficking of its proteolytic fragments J. Gen. Virol., July 1, 2004; 85(7): 1989 - 1994. [Abstract] [Full Text] [PDF]
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M. S. Mavinakere and A. M. Colberg-Poley Dual targeting of the human cytomegalovirus UL37 exon 1 protein during permissive infection J. Gen. Virol., February 1, 2004; 85(2): 323 - 329. [Abstract] [Full Text] [PDF]
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R. Adair, G. W. Liebisch, and A. M. Colberg-Poley Complex alternative processing of human cytomegalovirus UL37 pre-mRNA J. Gen. Virol., December 1, 2003; 84(12): 3353 - 3358. [Abstract] [Full Text] [PDF]
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Y. Su, R. Adair, C. N. Davis, N. L. DiFronzo, and A. M. Colberg-Poley Convergence of RNA cis Elements and Cellular Polyadenylation Factors in the Regulation of Human Cytomegalovirus UL37 Exon 1 Unspliced RNA Production J. Virol., December 1, 2003; 77(23): 12729 - 12741. [Abstract] [Full Text] [PDF]
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