A glycosylation-defective variant of the ecotropic murine retrovirus receptor is expressed in rat XC cells.
The XC rat sarcoma cell line undergoes extensive cell-to-cell fusion (syncytium formation) after infection with ecotropic murine leukemia viruses (MLVs) and is frequently used to titrate these viruses. This cell line is unique in its response to the ecotropic MLV envelope (Env) protein in that it undergoes syncytium formation with cells expressing Env protein containing R peptide (R+ Env), which is known to suppress the fusogenic potential of the Env protein in other susceptible cells. To assess whether this property of the XC cell line arises from differences in its ecotropic MLV receptor, CAT1, we isolated CAT1 cDNA clones from XC cells. A variant CAT1 (xcCAT1) was found together with the wild-type rat CAT1 (rCAT1). xcCAT1 cDNA encodes a protein with a single amino acid change that destroys a conserved N-linked glycosylation site proximal to the Env-binding motif. We found that xcCAT1 expressed in Chinese hamster ovary (CHO) cells undergoes less glycosylation than rCAT1 and that the expression of xcCAT1 rendered the CHO cells more susceptible to infection with Moloney MLV. Thus, N-glycosylation negatively regulates the receptor activity of rCAT1. This is supported by the observation that treatment of rat F10 cells with the N-glycosylation inhibitor tunicamycin enhanced their susceptibility to Mo-MLV. However, xcCAT1-expressing CHO cells did not fuse with 293T cells expressing R+ Env, indicating that xcCAT1 expression is not sufficient to induce the XC cell-specific characteristic of forming syncytia in response to R+ Env.
Kubo
Yoshinao
Y
Molecular Cell Science Laboratory, RIKEN Tsukuba Institute, Tsukuba, Ibaraki, Japan. yoshinao@postman.riken.go.jp
Ono
Takeshi
T
Ogura
Miki
M
Ishimoto
Akinori
A
Amanuma
Hiroshi
H
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D027201:Cationic Amino Acid Transporter 1 / Q000378:metabolism*; D006224:Cricetinae; D018076:DNA, Complementary / Q000302:isolation & purification; D015686:Gene Products, env / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000502:physiology*; D051381:Rats; D011991:Receptors, Virus / Q000378:metabolism*; D012509:Sarcoma / Q000821:virology*; D014407:Tumor Cells, Cultured
D027201:Cationic Amino Acid Transporter 1; D018076:DNA, Complementary; D015686:Gene Products, env; D011991:Receptors, Virus