The paramyxovirus SV5 small hydrophobic (SH) protein is not essential for virus growth in tissue culture cells.

The SH gene of the paramyxovirus SV5 is located between the genes for the glycoproteins, fusion protein (F) and hemagglutinin-neuraminidase (HN), and the SH gene encodes a small 44-residue hydrophobic integral membrane protein (SH). The SH protein is expressed in SV5-infected cells and is oriented in membranes with its N terminus in the cytoplasm. To study the function of the SH protein in the SV5 virus life cycle, the SH gene was deleted from the infectious cDNA clone of the SV5 genome. By using the recently developed reverse genetics system for SV5, it was found that an SH-deleted SV5 (rSV5DeltaSH) could be recovered, indicating the SH protein was not essential for virus viability in tissue culture. Analysis of properties of rSV5DeltaSH indicated that lack of expression of SH protein did not alter the expression level of the other virus proteins, the subcellular localization of F and HN, or fusion competency as measured by lipid mixing assays and a new content mixing assay that did not require the use of vaccinia virus. The growth rate, infectivity, and plaque size of rSV5 and rSV5DeltaSH were found to be very similar. Although SH is shown to be a component of purified virions by immunoblotting, examination of purified rSV5DeltaSH by electron microscopy did not show an altered morphology from SV5. Thus in tissue culture cells the lack of the SV5 SH protein does not confer a recognizable phenotype.

[1]  A. Roberts,et al.  Recovery of negative-strand RNA viruses from plasmid DNAs: a positive approach revitalizes a negative field. , 1998, Virology.

[2]  S S Whitehead,et al.  Respiratory syncytial virus (RSV) SH and G proteins are not essential for viral replication in vitro: clinical evaluation and molecular characterization of a cold-passaged, attenuated RSV subgroup B mutant. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[3]  B. Murphy,et al.  Recombinant respiratory syncytial virus from which the entire SH gene has been deleted grows efficiently in cell culture and exhibits site-specific attenuation in the respiratory tract of the mouse , 1997, Journal of virology.

[4]  R. Lamb,et al.  A glycine to alanine substitution in the paramyxovirus SV5 fusion peptide increases the initial rate of fusion. , 1997, Virology.

[5]  R. Lamb,et al.  Recovery of infectious SV5 from cloned DNA and expression of a foreign gene. , 1997, Virology.

[6]  T. Yoshida,et al.  Importance of the cysteine-rich carboxyl-terminal half of V protein for Sendai virus pathogenesis , 1997, Journal of virology.

[7]  G. Parks,et al.  Genome nucleotide lengths that are divisible by six are not essential but enhance replication of defective interfering RNAs of the paramyxovirus simian virus 5. , 1997, Virology.

[8]  R. Lamb,et al.  Influenza virus hemagglutinin and neuraminidase cytoplasmic tails control particle shape , 1997, The EMBO journal.

[9]  D. Garcin,et al.  Normal cellular replication of Sendai virus without the trans-frame, nonstructural V protein. , 1997, Virology.

[10]  Y. Nagai,et al.  The paramyxovirus, Sendai virus, V protein encodes a luxury function required for viral pathogenesis , 1997, The EMBO journal.

[11]  M. Billeter,et al.  Recombinant measles viruses defective for RNA editing and V protein synthesis are viable in cultured cells. , 1997, Virology.

[12]  A. Yamada,et al.  The mumps virus SH protein is a membrane protein and not essential for virus growth. , 1996, Virology.

[13]  R. Lamb,et al.  Truncation of the COOH-terminal region of the paramyxovirus SV5 fusion protein leads to hemifusion but not complete fusion , 1996, The Journal of cell biology.

[14]  R. Lamb,et al.  Influenza B virus NB glycoprotein is a component of the virion. , 1996, Virology.

[15]  M. Whitt,et al.  Normal replication of vesicular stomatitis virus without C proteins. , 1996, Virology.

[16]  R. Lamb,et al.  Quantitative measurement of paramyxovirus fusion: differences in requirements of glycoproteins between simian virus 5 and human parainfluenza virus 3 or Newcastle disease virus , 1995, Journal of virology.

[17]  W. Mcallister,et al.  Phage RNA polymerase vectors that allow efficient gene expression in both prokaryotic and eukaryotic cells. , 1995, Gene.

[18]  G. Sutter,et al.  Non‐replicating vaccinia vector efficiently expresses bacteriophage T7 RNA polymerase , 1995, FEBS letters.

[19]  B. Moss,et al.  Replication-deficient vaccinia virus encoding bacteriophage T7 RNA polymerase for transient gene expression in mammalian cells. , 1995, Virology.

[20]  R. Lamb,et al.  The paramyxovirus SV5 V protein binds two atoms of zinc and is a structural component of virions. , 1995, Virology.

[21]  E. Schreier,et al.  Differentiation of vaccine and wild mumps viruses by polymerase chain reaction and nucleotide sequencing of the SH gene: Brief report , 1995, Journal of medical virology.

[22]  C. Broder,et al.  Fusogenic mechanisms of enveloped-virus glycoproteins analyzed by a novel recombinant vaccinia virus-based assay quantitating cell fusion-dependent reporter gene activation , 1994, Journal of virology.

[23]  K. Conzelmann,et al.  Infectious rabies viruses from cloned cDNA. , 1994, The EMBO journal.

[24]  Y. Tanaka,et al.  Analysis of respiratory syncytial virus F, G, and SH proteins in cell fusion. , 1994, Virology.

[25]  R. Lamb.  Paramyxovirus fusion: a hypothesis for changes. , 1993, Virology.

[26]  L. Roux,et al.  The rule of six, a basic feature for efficient replication of Sendai virus defective interfering RNA , 1993, Journal of virology.

[27]  A. Heath,et al.  The Jeryl Lynn vaccine strain of mumps virus is a mixture of two distinct isolates. , 1993, The Journal of general virology.

[28]  R. Lamb,et al.  Studies on the fusion peptide of a paramyxovirus fusion glycoprotein: roles of conserved residues in cell fusion , 1992, Journal of virology.

[29]  R. Lamb,et al.  Molecular cloning of the NP and L genes of simian virus 5: identification of highly conserved domains in paramyxovirus NP and L proteins. , 1992, Virus research.

[30]  M. Tsurudome,et al.  Molecular relationships between human parainfluenza virus type 2, and simian viruses 41 and 5: determination of nucleoprotein gene sequences of simian viruses 41 and 5. , 1991, The Journal of general virology.

[31]  A. Yamada,et al.  Variations of nucleotide sequences and transcription of the SH gene among mumps virus strains. , 1991, Virology.

[32]  Y. Ito,et al.  Complete nucleotide sequence of the matrix gene of human parainfluenza type 2 virus and expression of the M protein in bacteria. , 1990, Virology.

[33]  W. Mcallister,et al.  Synthesis of functional mRNA in mammalian cells by bacteriophage T3 RNA polymerase , 1990, Molecular and cellular biology.

[34]  Y. Ito,et al.  Molecular cloning and sequence analysis of human parainfluenza type 4A virus HN gene: its irregularities on structure and activities. , 1990, Virology.

[35]  R. Lamb,et al.  Intracellular maturation and transport of the SV5 type II glycoprotein hemagglutinin-neuraminidase: specific and transient association with GRP78-BiP in the endoplasmic reticulum and extensive internalization from the cell surface , 1989, The Journal of cell biology.

[36]  P. Collins,et al.  The 1A protein of respiratory syncytial virus is an integral membrane protein present as multiple, structurally distinct species , 1989, Journal of virology.

[37]  D. P. Sarkar,et al.  Kinetics of pH-dependent fusion between 3T3 fibroblasts expressing influenza hemagglutinin and red blood cells. Measurement by dequenching of fluorescence. , 1989, The Journal of biological chemistry.

[38]  E. Norrby,et al.  mRNA sequence and deduced amino acid sequence of the mumps virus small hydrophobic protein gene , 1989, Journal of virology.

[39]  Sheila M. Thomas,et al.  Two mRNAs that differ by two nontemplated nucleotides encode the amino coterminal proteins P and V of the paramyxovirus SV5 , 1988, Cell.

[40]  R. Lamb,et al.  Influenza A virus M2 protein: monoclonal antibody restriction of virus growth and detection of M2 in virions , 1988, Journal of virology.

[41]  A. Yamada,et al.  Molecular cloning and sequence analysis of the mumps virus gene encoding the P protein: mumps virus P gene is monocistronic. , 1988, The Journal of general virology.

[42]  R. Lamb,et al.  Cell surface expression and orientation in membranes of the 44-amino-acid SH protein of simian virus 5 , 1988, Journal of virology.

[43]  R. Randall,et al.  Isolation and characterization of monoclonal antibodies to simian virus 5 and their use in revealing antigenic differences between human, canine and simian isolates. , 1987, The Journal of general virology.

[44]  R. Lamb,et al.  Identification and predicted sequence of a previously unrecognized small hydrophobic protein, SH, of the paramyxovirus simian virus 5 , 1985, Journal of virology.

[45]  R. Lamb,et al.  Hemagglutinin-neuraminidase protein of the paramyxovirus simian virus 5: nucleotide sequence of the mRNA predicts an N-terminal membrane anchor , 1985, Journal of virology.

[46]  P. Collins,et al.  The 1A protein gene of human respiratory syncytial virus: nucleotide sequence of the mRNA and a related polycistronic transcript. , 1985, Virology.

[47]  R. Lamb,et al.  Analysis and gene assignment of mRNAs of a paramyxovirus, simian virus 5. , 1984, Virology.

[48]  W. Russell,et al.  Does simian virus 5 infect humans? , 1984, The Journal of general virology.

[49]  W. N. Burnette,et al.  "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. , 1981, Analytical biochemistry.

[50]  R. Lamb,et al.  Synthesis of influenza virus proteins in infected cells: translation of viral polypeptides, including three P polypeptides, from RNA produced by primary transcription. , 1976, Virology.

[51]  P. Choppin.  MULTIPLICATION OF A MYXOVIRUS (SV5) WITH MINIMAL CYTOPATHIC EFFECTS AND WITHOUT INTERFERENCE. , 1964, Virology.

[52]  J. W. Smith,et al.  New viral agents recovered from tissue cultures of monkey kidney cells. I. Origin and properties of cytopathogenic agents S.V.1, S.V.2, S.V.4, S.V.5, S.V.6, S.V.11, S.V.12 and S.V.15. , 1956, American journal of hygiene.

[53]  R. Lamb,et al.  Orthomyxoviridae: The Viruses and Their Replication. , 1996 .

[54]  R. Lamb,et al.  The molecular biology of influenza viruses and paramyxoviruses , 1993 .

[55]  A. Davison,et al.  Molecular virology: a practical approach , 1993 .