Internal deletions of IE2 86 and loss of the late IE2 60 and IE2 40 proteins encoded by human cytomegalovirus affect the levels of UL84 protein but not the amount of UL84 mRNA or the loading and distribution of the mRNA on polysomes.
The major immediate-early (IE) region of human cytomegalovirus encodes two IE proteins, IE1 72 and IE2 86, that are translated from alternatively spliced transcripts that differ in their 3' ends. Two other proteins that correspond to the C-terminal region of IE2 86, IE2 60 and IE2 40, are expressed at late times. In this study, we used IE2 mutant viruses to examine the mechanism by which IE2 86, IE2 60, and IE2 40 affect the expression of a viral DNA replication factor, UL84. Deletion of amino acids (aa) 136 to 290 of IE2 86 results in a significant decrease in UL84 protein during the infection. This loss of UL84 is both proteasome and calpain independent, and the stability of the protein in the context of infection with the mutant remains unaffected. The RNA for UL84 is expressed to normal levels in the mutant virus-infected cells, as are the RNAs for two other proteins encoded by this region, UL85 and UL86. Moreover, nuclear-to-cytoplasmic transport and the distribution of the UL84 mRNA on polysomes are unaffected. A region between aa 290 and 369 of IE2 86 contributes to the UL84-IE2 86 interaction in vivo and in vitro. IE2 86, IE2 60, and IE2 40 are each able to interact with UL84 in the mutant-infected cells, suggesting that these interactions may be important for the roles of UL84 and the IE2 proteins. Thus, these data have defined the contribution of IE2 86, IE2 60, and IE2 40 to the efficient expression of UL84 throughout the infection.
Sanders
Rebecca L
RL
Skaggs School of Pharmacy and Pharmaceutical Sciences, Room 3254, Mail Code 0712, 9500 Gilman Drive, University of California, San Diego, La Jolla, CA 92093-0712, USA. dspector@ucsd.edu
Del Rosario
Christia J
CJ
White
Elizabeth A
EA
Spector
Deborah H
DH
D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics* / Q000502:physiology; D017353:Gene Deletion; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011132:Polyribosomes / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis*; D012367:RNA, Viral / Q000096:biosynthesis*; D017384:Sequence Deletion; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism*; D014764:Viral Proteins / Q000096:biosynthesis*
D036022:Capsid Proteins; C069186:IE2 protein, Cytomegalovirus; D017874:Immediate-Early Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D015534:Trans-Activators; D014764:Viral Proteins; C443660:major capsid protein, Cytomegalovirus; C072218:UL84 protein, Cytomegalovirus
J Virol. 1983 Apr;46(1):1-14
6298447
J Virol. 2008 Jul;82(14):7059-77
18463148
Adv Virus Res. 1999;54:61-128
10547675
J Virol. 1990 Dec;64(12):6154-65
2173785
Virology. 1993 Apr;193(2):642-52
7681609
J Virol. 2004 Oct;78(19):10360-9
15367602
J Virol. 1993 Oct;67(10):6201-14
8396676
J Virol. 1985 Dec;56(3):665-75
2999423
J Virol. 2006 Apr;80(8):3872-83
16571804
J Virol. 1994 Nov;68(11):7549-53
7933141
J Virol. 2007 Jul;81(13):7077-85
17459920
J Virol. 2004 Sep;78(17):9203-14
15308715
J Biol Chem. 2005 Mar 25;280(12):11955-60
15778228
Gene. 1998 Mar 27;210(1):25-36
9524210
J Virol. 1995 Oct;69(10):6533-40
7666555
J Virol. 2005 Jun;79(12):7438-52
15919900
J Virol. 1991 Feb;65(2):887-96
1846203
Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):707-11
8380646
J Virol. 2001 Feb;75(4):1870-8
11160686
J Virol. 1991 Jul;65(7):3839-52
1645794
J Virol. 1995 Dec;69(12):7612-21
7494269
J Virol. 2001 Apr;75(8):3859-72
11264375
Biochem Biophys Res Commun. 1988 Jun 30;153(3):1201-8
2839170
Proc Natl Acad Sci U S A. 2003 Oct 14;100(21):12396-401
14519856
J Virol. 1998 Jul;72(7):5481-92
9621004
J Virol. 1984 Jan;49(1):190-9
6317889
J Virol. 1999 Oct;73(10):8320-9
10482582
J Virol. 1990 Apr;64(4):1556-65
2157043
J Virol. 1989 Jun;63(6):2699-708
2542583
J Virol. 1997 Sep;71(9):7048-60
9261435
J Virol. 2003 Apr;77(8):4760-72
12663783
J Virol. 1992 Feb;66(2):1098-108
1309892
J Virol. 1994 Sep;68(9):5613-22
8057441
J Med Virol. 2002 May;67(1):33-40
11920815
J Biol Chem. 1987 Oct 25;262(30):14806-14
2444591
J Virol. 1993 Aug;67(8):4981-91
8392623
J Virol. 2004 Nov;78(21):11664-77
15479808
J Virol. 1993 Dec;67(12):6979-88
8230421
J Gen Virol. 1994 Sep;75 ( Pt 9):2337-48
8077932
J Virol. 1991 Nov;65(11):6301-6
1656096
Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11321-6
8876134
Virology. 1998 Dec 5;252(1):235-57
9875333
J Virol. 2000 Mar;74(6):2510-24
10684265
J Virol. 1994 Oct;68(10):6223-31
8083962
Virology. 1993 Aug;195(2):786-92
8337845
J Virol. 2005 Feb;79(3):1428-37
15650169
J Virol. 2004 Feb;78(4):1817-30
14747546
Mol Cell Biol. 1993 Feb;13(2):1238-50
8423789
J Virol. 2007 Mar;81(6):2573-83
17202222
J Virol. 2005 Sep;79(17):11467-75
16103197
J Virol. 2007 Jun;81(11):5807-18
17376893
J Virol. 2002 Mar;76(6):2973-89
11861863
J Virol. 2005 Jan;79(1):225-33
15596818
J Virol. 1998 Jan;72(1):366-79
9420235
J Virol. 2002 May;76(9):4441-55
11932411
J Virol. 1991 Feb;65(2):897-903
1846204
J Virol. 1990 Apr;64(4):1498-506
2157038
Nucleic Acids Res. 1994 Aug 25;22(16):3331-8
8078768
J Virol. 1994 Feb;68(2):720-9
8289376
FEBS Lett. 2003 Dec 4;555(2):322-8
14644436
J Gen Virol. 2004 Aug;85(Pt 8):2149-2154
15269353
J Virol. 2004 Jul;78(14):7803-12
15220454
J Virol. 1996 Nov;70(11):7398-413
8892858
J Virol. 1990 Oct;64(10):5167-72
2168993