title: Bovine leukemia virus gp30 transmembrane (TM) protein is not tyrosine phosphorylated: examining potential interactions with host tyrosine-mediated signaling.
abstract: Bovine leukemia virus (BLV) causes persistent lymphocytosis, a preneoplastic, polyclonal expansion of B lymphocytes. The expansion increases viral transmission to new hosts, but the mechanisms of this expansion have not been determined. We hypothesized that BLV infection contributes to B-cell expansion by signaling initiated via viral transmembrane protein motifs undergoing tyrosine phosphorylation. Viral mimicry of host cell proteins is a well-demonstrated mechanism by which viruses may increase propagation or decrease recognition by the host immune system. The cytoplasmic tail of BLV transmembrane protein gp30 (TM) has multiple areas of homology to motifs of host cell signaling proteins, including two immunoreceptor tyrosine-based activation motifs (ITAMs) and two immunoreceptor tyrosine-based inhibition motifs (ITIMs), which are homologous to B-cell receptor and inhibitory co-receptor motifs. Signaling by these motifs in B cells typically relies on tyrosine phosphorylation, followed by interactions with Src-homology-2 (SH2) domains of nonreceptor protein tyrosine kinases or phosphatases. Phosphorylation of tyrosine residues in the cytoplasmic tail of TM was tested in four systems including ex vivo cultured peripheral blood mononuclear cells from BLV infected cows, BLV-expressing fetal lamb kidney cell and bat lung cell lines, and DT40 B cells transfected with a fusion of mouse extracellular CD8alpha and cytoplasmic TM. No phosphorylation of TM was detected in our experiments in any of the cell types utilized, or with various stimulation methods. Detection was attempted by immunoblotting for phosphotyrosines, or by metabolic labeling of cells. Thus BLV TM is not likely to modify host signal pathways through interactions between phosphorylated tyrosines of the ITAM or ITIM motifs and host-cell tyrosine kinases or phosphatases.
authors.affiliation: Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040 USA.
authors.forename: Valerie T
authors.initials: VT
authors.lastname: Hamilton
authors.forename: Diana M
authors.initials: DM
authors.lastname: Stone
authors.forename: Suzanne M
authors.initials: SM
authors.lastname: Pritchard
authors.forename: Glenn H
authors.initials: GH
authors.lastname: Cantor
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D002417:Cattle; D002460:Cell Line; D016583:Enzootic Bovine Leukosis / Q000821:virology; D001909:Leukemia Virus, Bovine / Q000472:pathogenicity*; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017459:Receptors, Amino Acid / Q000737:chemistry / Q000378:metabolism; D011947:Receptors, Antigen, B-Cell / Q000737:chemistry / Q000378:metabolism; D011971:Receptors, Immunologic / Q000737:chemistry / Q000378:metabolism; D015682:Retroviridae Proteins, Oncogenic / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction; D014443:Tyrosine / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D017459:Receptors, Amino Acid; D011947:Receptors, Antigen, B-Cell; D011971:Receptors, Immunologic; D015682:Retroviridae Proteins, Oncogenic; D014759:Viral Envelope Proteins; C060711:gp30 envelope transmembrane protein, Bovine leukemia virus; C087659:tyrosine receptor; D014443:Tyrosine