Transcription inhibition site on the M protein of vesicular stomatitis virus located by marker rescue of mutant tsO23(III) with M-gene expression vectors

Abstract

The matrix (M) protein of vesicular stomatitis virus serves as an endogenous inhibitor of viral transcription, a function missing or deficient in M proteins of temperature-sensitive (ts) mutants assigned to complementation group III. Previous studies with mutant tsO23(III) and vaccinia virus M-gene expression vectors revealed that the temperature-sensitive phenotype is due to a mutation leading to substitution of phenylalanine for leucine at amino acid III, whereas loss of the major antigenic determinant (epitope 1) of the mutant M protein results from the substitution of glutamic acid for the wild-type amino acid glycine at position 21 (Y.

Monoclonal Antibody #3‐9‐16 Recognizes One of the Two Isoforms of Rabies Virus Matrix Protein That Exposes Its N‐Terminus on the Virion Surface

“…The M protein of vesicular stomatitis virus (VSV), another major member of the Rhabdoviridae, was extensively studied, and is now known to work multifunctionally in the virus replication cycle, such as the regulation of viral RNA synthesis, stabilization of viral glycoprotein and virion formation with G and RNP at the budding site (1,2,4,15,(19)(20)(21).…”

Inducible and conditional inhibition of human immunodeficiency virus proviral expression by vesicular stomatitis virus matrix protein

“…This indicated that M protein may be toxic to the cell and/or to vaccinia virus. Transient expression of M protein from plasmid DNA by T7 RNA polymerase encoded by a recombinant vaccinia virus, however, was efficient and allowed to complement M protein mutants of VSV (18,19). A recombinant vaccinia virus that expresses M protein of the closely related rabies virus has been isolated (14).…”

Membrane vesiculation function and exocytosis of wild-type and mutant matrix proteins of vesicular stomatitis virus

“…This membrane-budding function is also shown to be restricted by a specific site mutation in the M-gene cDNA of a VSV temperature-sensitive mutant, in which phenylalanine is substituted for leucine at amino acid 111 of the M protein. This mutation previously had been shown to be responsible for restricted production of virions at a nonpermissive temperature (11), which can also be rescued by a coexpressed revertant (10).…”

“…The wild-type (wt) M gene studied here is a cDNA clone coding for all 229 amino acids of the VSV M protein; it was obtained from mRNA of VSV-Indiana (Orsay strain) and was cloned in this laboratory into a vector designated pYL-OM79, flanked by promoter and terminator sequences for bacteriophage T7 RNA polymerase (10). We had also created deletion mutants of the M gene by restriction enzyme cleavage or by PCR which gave rise to truncated proteins in which the first 50 amino acids were deleted or amino acids 75 to 229 were deleted, as well as a double-deletion mutant missing amino acids 1 to 50 and 75 to 106 (20).…”

“…We had also created deletion mutants of the M gene by restriction enzyme cleavage or by PCR which gave rise to truncated proteins in which the first 50 amino acids were deleted or amino acids 75 to 229 were deleted, as well as a double-deletion mutant missing amino acids 1 to 50 and 75 to 106 (20). In addition, the cDNA of the M gene of the temperature-sensitive mutant tsO23 had also been cloned in the same expression vector (10); the recombinant tsO23 M gene gave rise to a protein with amino acid substitutions at residues 21, 111, and 227 (11). Also available from earlier studies were mutant clones of the wt M gene which gave rise to proteins with single-amino-acid substitutions at residues 21, 111, and 227 identical to those of the tsO23 mutations, as well as revertants in the tsO23 M gene expressing proteins with individual revertants at amino acids 21, 111, and 227 (10).…”