GRB2 interaction with the ecotropic murine leukemia virus receptor, mCAT-1, controls virus entry and is stimulated by virus binding

Zeming Chen, Andrey A. Kolokoltsov, Jia Wang, Shramika Adhikary, Marta Lorinczi, Lisa Elferink, Robert A. Davey

Research output: Contribution to journalArticle

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Abstract

For retroviruses such as HIV-1 and murine leukemia virus (MLV), active receptor recruitment and trafficking occur during viral entry. However, the underlying mechanisms and cellular factors involved in the process are largely uncharacterized. The viral receptor for ecotropic MLV (eMLV), a classical model for retrovirus infection mechanisms and pathogenesis, is mouse cationic amino acid transporter 1 (mCAT-1). Growth factor receptor-bound protein 2 (GRB2) is an adaptor protein that has been shown to couple cell surface receptors, such as epidermal growth factor receptor (EGFR) and hepatocyte growth factor receptor, to intracellular signaling events. Here we examined if GRB2 could also play a role in controlling infection by retroviruses by affecting receptor function. The GRB2 RNA interference (RNAi)-mediated suppression of endogenous GRB2 resulted in a consistent and significant reduction of virus binding and membrane fusion. The binding between eMLV and cells promoted increased GRB2-mCAT-1 interactions, as detected by immunoprecipitation. Consistently, the increased colocalization of GRB2 and mCAT-1 signals was detected by confocal microscopy. This association was time dependent and paralleled the kinetics of cell-virus membrane fusion. Interestingly, unlike the canonical binding pattern seen for GRB2 and growth factor receptors, GRB2-mCAT-1 binding does not depend on the GRB2-SH2 domain-mediated recognition of tyrosine phosphorylation on the receptor. The inhibition of endogenous GRB2 led to a reduction in surface levels of mCAT-1, which was detected by immunoprecipitation and by a direct binding assay using a recombinant MLV envelope protein receptor binding domain (RBD). Consistent with this observation, the expression of a dominant negative GRB2 mutant (R86K) resulted in the sequestration of mCAT-1 from the cell surface into intracellular vesicles. Taken together, these findings suggest a novel role for GRB2 in ecotropic MLV entry and infection by facilitating mCAT-1 trafficking.

Original languageEnglish (US)
Pages (from-to)1421-1432
Number of pages12
JournalJournal of Virology
Volume86
Issue number3
DOIs
StatePublished - Feb 2012

Fingerprint

Cationic Amino Acid Transporter 1
GRB2 Adaptor Protein
Murine leukemia virus
amino acid transporters
Virus Attachment
Virus Internalization
growth factors
viruses
receptors
mice
Murine Leukemia Viruses
proteins
Retroviridae Infections
Retroviridae
Immunoprecipitation
Proto-Oncogene Proteins c-met
Viral Envelope Proteins
Virus Receptors
ecotropic murine leukemia virus receptor
virus receptors

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

GRB2 interaction with the ecotropic murine leukemia virus receptor, mCAT-1, controls virus entry and is stimulated by virus binding. / Chen, Zeming; Kolokoltsov, Andrey A.; Wang, Jia; Adhikary, Shramika; Lorinczi, Marta; Elferink, Lisa; Davey, Robert A.

In: Journal of Virology, Vol. 86, No. 3, 02.2012, p. 1421-1432.

Research output: Contribution to journalArticle

Chen, Zeming ; Kolokoltsov, Andrey A. ; Wang, Jia ; Adhikary, Shramika ; Lorinczi, Marta ; Elferink, Lisa ; Davey, Robert A. / GRB2 interaction with the ecotropic murine leukemia virus receptor, mCAT-1, controls virus entry and is stimulated by virus binding. In: Journal of Virology. 2012 ; Vol. 86, No. 3. pp. 1421-1432.
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