Murine Cytomegalovirus Protein pM49 Interacts with pM95 and Is Critical for Viral Late Gene Expression.

Publisher: American Society For Microbiology Country of Publication: United States NLM ID: 0113724 Publication Model: Electronic-Print Cited Medium: Internet ISSN: 1098-5514 (Electronic) Linking ISSN: 0022538X NLM ISO Abbreviation: J Virol Subsets: MEDLINE

Publication: Washington Dc : American Society For Microbiology
Original Publication: Baltimore, American Society for Microbiology.

Gene Expression Regulation, Viral* ; Mutation*; DNA, Viral/*biosynthesis ; Herpesviridae Infections/*metabolism ; Muromegalovirus/*metabolism ; Viral Proteins/*metabolism; Animals ; Cell Line ; DNA, Viral/genetics ; Herpesviridae Infections/genetics ; Mice ; Muromegalovirus/genetics ; Viral Proteins/genetics

Late gene expression of betaherpesviruses and gammaherpesviruses is tightly controlled by virus-encoded transactivation factors (vTFs). We recently proved that the 6 vTFs of murine cytomegalovirus (MCMV) form a complex to regulate late gene transcription. pM49, one of the vTFs that has not been studied before, was identified to be a component of the complex that interacts with pM95. In this study, we began to investigate the potential role of pM49 in viral late gene expression. A recombinant MCMV expressing C-terminal FLAG-tagged pM49 was constructed to study the expression kinetics and localization of pM49. pM49 was expressed at the late time of virus infection. Inhibition of viral DNA synthesis by phosphonate sodium phosphonic acid (PAA) abolished pM49 expression, indicating that it is a late protein. pM49 colocalized with pM44 at the viral replication compartment, similarly to other viral vTFs that have been reported. Mutant virus lacking full-length pM49 expression failed to express viral late genes, leading to nonproductive infection. The expression of immediate early and early genes was not affected, and viral DNA synthesis was only minimally affected during pM49-deficient virus infection. All of these data support the role of pM49 in viral late gene expression. After a series of mutagenesis analyses, two key residues, K325 and C326, were identified as required for pM49-pM95 interaction. Cells expressing pM49 with either single mutation of these two residues failed to rescue the late gene expression and support the replication of pM49-deficient virus. Our results indicated that pM49-pM95 interaction is essential for viral late gene expression. IMPORTANCE Cytomegalovirus (CMV) infections result in morbidity and mortality in immunocompromised individuals, and the virus is also a major cause of birth defects in newborns. Currently, because of the unavailability of vaccines against this virus and restricted antiviral therapies with low toxicity, as well as the emergency of resistant strain of this virus, the understanding of viral late gene regulation may provide clues to study new antiviral drugs or vaccines. In this study, we report that MCMV protein pM49 is critical for viral late gene transcription, based on its interaction with pM95. This finding reveals the important role of pM49-pM95 interaction in the regulation of viral late gene expression and that it could be a future potential target for therapeutic intervention in CMV diseases.
(Copyright © 2020 American Society for Microbiology.)

J Virol. 2014 Jan;88(1):120-30. (PMID: 24131715)
Virology. 1995 May 10;209(1):236-41. (PMID: 7747475)
Clin Microbiol Infect. 2007 Oct;13(10):953-63. (PMID: 17803749)
J Gen Virol. 2017 Feb;98(2):242-250. (PMID: 27926822)
J Virol. 2015 Mar;89(6):3049-61. (PMID: 25552713)
Hum Gene Ther. 1996 Aug 1;7(12):1405-13. (PMID: 8844199)
J Virol. 2020 Jan 6;94(2):. (PMID: 31578296)
Med Microbiol Immunol. 2008 Jun;197(2):233-40. (PMID: 18239940)
J Virol. 2004 Jun;78(12):6610-20. (PMID: 15163752)
J Virol. 1998 Oct;72(10):8338-43. (PMID: 9733880)
J Virol. 2014 Nov;88(21):12825-38. (PMID: 25165108)
Methods Mol Biol. 2010;634:421-30. (PMID: 20677001)
J Virol. 2013 Aug;87(15):8651-64. (PMID: 23720731)
Virology. 1988 Dec;167(2):477-84. (PMID: 2849236)
Biotechniques. 2006 Feb;40(2):191-7. (PMID: 16526409)
PLoS Pathog. 2016 Jun 27;12(6):e1005718. (PMID: 27348612)
J Virol. 2012 Jun;86(11):6023-32. (PMID: 22457524)
J Virol. 1997 Feb;71(2):1246-55. (PMID: 8995648)
J Virol. 1996 Mar;70(3):1855-62. (PMID: 8627710)
J Virol. 2011 Sep;85(17):9103-13. (PMID: 21715486)
J Virol. 2003 Mar;77(6):3602-14. (PMID: 12610136)
J Virol. 2014 Jan;88(1):131-42. (PMID: 24131717)
J Virol. 2009 Mar;83(5):2265-73. (PMID: 19091863)
J Virol. 2018 Aug 29;92(18):. (PMID: 29997217)
Sci Rep. 2016 May 18;6:26167. (PMID: 27188239)
J Virol. 2008 Apr;82(7):3452-65. (PMID: 18216115)
PLoS Pathog. 2014 Aug 28;10(8):e1004350. (PMID: 25166009)
J Virol. 2012 Jun;86(12):6712-23. (PMID: 22496230)
Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):11137-42. (PMID: 8855322)
Microbes Infect. 2003 Nov;5(13):1263-77. (PMID: 14623023)
PLoS Pathog. 2019 Aug 28;15(8):e1007980. (PMID: 31461506)
J Virol. 2015 Oct 14;90(1):599-604. (PMID: 26468530)
Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):3937-42. (PMID: 9520471)
Proc Natl Acad Sci U S A. 1975 Apr;72(4):1276-80. (PMID: 165503)
J Virol. 1999 Aug;73(8):7056-60. (PMID: 10400809)
J Virol. 2013 Aug;87(16):9135-47. (PMID: 23760242)
Genes Dev. 1991 Dec;5(12B):2375-85. (PMID: 1752433)
J Virol. 1996 Dec;70(12):8833-49. (PMID: 8971012)
J Virol. 2006 Oct;80(19):9730-40. (PMID: 16973577)
Front Microbiol. 2016 Jun 06;7:869. (PMID: 27375590)
Lancet. 2004 Jun 26;363(9427):2116-21. (PMID: 15220032)
Proc Natl Acad Sci U S A. 1974 Nov;71(11):4322-6. (PMID: 4373710)
J Virol. 2020 Jan 6;94(2):. (PMID: 31694948)

Keywords: cytomegalovirus; late gene; pM49; pM95; transactivation factors

0 (DNA, Viral)
0 (Viral Proteins)

Date Created: 20200104 Date Completed: 20200902 Latest Revision: 20200902

20240513

PMC7158740

10.1128/JVI.01956-19

31896598