Rhesus rhadinovirus infection of rhesus fibroblasts occurs through clathrin-mediated endocytosis.
Rhesus rhadinovirus (RRV) is a gammaherpesvirus closely related to Kaposi's sarcoma-associated herpesvirus (KSHV), an oncogenic virus linked to the development of Kaposi's sarcoma and several other lymphoproliferative diseases, including primary effusion lymphoma and multicentric Castleman's disease. RRV naturally infects rhesus macaques and induces lymphoproliferative diseases under experimental conditions, making it an excellent model for the study of KSHV. Unlike KSHV, which grows poorly in cell culture, RRV replicates efficiently in rhesus fibroblasts (RFs). In this study, we have characterized the entry pathway of RRV in RFs. Using a luciferase-expressing recombinant RRV (RRV-luciferase), we show that the infectivity of RRV is reduced by inhibitors of endosomal acidification. RRV infectivity is also reduced by inhibitors of clathrin-mediated but not caveola-mediated endocytosis, indicating that RRV enters into RFs via clathrin-mediated endocytosis. Using a red fluorescent protein (RFP)-expressing recombinant RRV (RRV-RFP), we show that RRV particles are colocalized with markers of endocytosis (early endosome antigen 1) and clathrin-mediated endocytosis (clathrin heavy chain) during entry into RFs. RRV particles are also colocalized with transferrin, which enters cells by clathrin-mediated endocytosis, but not with cholera toxin B, which enters cells by caveola-mediated endocytosis. Inhibition of clathrin-mediated endocytosis with a dominant-negative construct of EPS15, an essential component of clathrin-coated pits, blocked the entry of RRV into RFs. Together, these results indicate that RRV entry into RFs is mediated by clathrin-mediated endocytosis.
Zhang
Wei
W
The University of Texas Health Science Center, San Antonio, TX 78229, USA.
Zhou
Fuchun
F
Greene
Whitney
W
Gao
Shou-Jiang
SJ
D000818:Animals; D002478:Cells, Cultured; D002966:Clathrin / Q000378:metabolism*; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism / Q000821:virology; D005347:Fibroblasts / Q000378:metabolism* / Q000821:virology; D006566:Herpesviridae Infections / Q000378:metabolism* / Q000821:virology; D019288:Herpesvirus 8, Human / Q000502:physiology; D006801:Humans; D008253:Macaca mulatta / Q000821:virology; D018144:Rhadinovirus / Q000502:physiology*
D002966:Clathrin
Proc Natl Acad Sci U S A. 1987 Aug;84(15):5454-8
3037552
J Virol. 2008 May;82(10):4793-806
18337571
Cell. 2009 May 1;137(3):433-44
19410541
J Virol. 2007 Aug;81(15):7825-32
17459936
J Virol. 2003 Oct;77(19):10179-85
12970403
Virology. 2010 May 10;400(2):175-86
20172576
Proc Natl Acad Sci U S A. 1988 Nov;85(21):7972-6
2973058
Curr Biol. 2001 Nov 27;11(23):1880-4
11728312
Biochemistry. 1995 Mar 14;34(10):3455-60
7880839
J Virol. 1999 Apr;73(4):3040-53
10074154
PLoS Pathog. 2009 Jul;5(7):e1000512
19593382
Cell. 1992 Feb 21;68(4):673-82
1739974
J Virol. 2009 May;83(10):4895-911
19279100
J Virol. 2008 Feb;82(3):1570-80
18045938
Cell. 2002 Feb 8;108(3):407-19
11853674
Science. 2006 Mar 31;311(5769):1921-4
16574866
J Cell Biol. 1993 Dec;123(5):1107-17
8245121
Biotechniques. 2006 Feb;40(2):191-7
16526409
Blood. 2008 Nov 15;112(10):4227-34
18757778
J Exp Med. 1999 Sep 20;190(6):827-40
10499921
Nat Rev Mol Cell Biol. 2009 Sep;10(9):597-608
19696797
J Cell Biol. 1998 Mar 9;140(5):1055-62
9490719
Cancer Treat Res. 2007;133:69-127
17672038
J Virol. 2000 Apr;74(7):3388-98
10708456
J Virol. 2003 May;77(9):5324-32
12692234
J Cell Sci. 2004 Mar 15;117(Pt 8):1421-30
14996913
J Cell Biol. 1989 Feb;108(2):389-400
2563728
J Virol. 1999 Dec;73(12):10320-8
10559350
J Virol. 1997 Dec;71(12):9764-9
9371642
Biochem J. 2008 Jun 15;412(3):415-23
18498251
AAPS J. 2009 Mar;11(1):65-77
19194803
J Virol. 2010 Mar;84(6):2871-80
20071577
J Virol. 2005 Jun;79(11):6655-63
15890903
Cell. 2006 Feb 24;124(4):729-40
16497584
J Med Microbiol. 2000 Apr;49(4):313-318
10755624
J Gen Virol. 2002 Jul;83(Pt 7):1535-1545
12075072
Cell. 2009 May 1;137(3):402-4
19410537
Curr Top Microbiol Immunol. 2007;312:43-69
17089793
J Virol. 2002 Jun;76(12):6185-96
12021352
J Virol. 2003 Jul;77(14):7978-90
12829837
J Cell Sci. 1999 May;112 ( Pt 9):1303-11
10194409