Truncation of the COOH-terminal region of the paramyxovirus SV5 fusion protein leads to hemifusion but not complete fusion

被引:65
作者
Bagai, S
Lamb, RA
机构
[1] NORTHWESTERN UNIV, DEPT BIOCHEM MOL BIOL & CELL BIOL, EVANSTON, IL 60208 USA
[2] NORTHWESTERN UNIV, HOWARD HUGHES MED INST, EVANSTON, IL 60208 USA
关键词
D O I
10.1083/jcb.135.1.73
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The role of the simian virus 5 (SV5) fusion (F) protein 20 residue COOH-terminal region, thought to represent the cytoplasmic tail, in fusion activity was examined by constructing a series of COOH-terminal truncation mutants. When the altered F proteins were expressed in eukaryotic cells, by using the vaccinia virus-T7 transient expression system, all the F proteins exhibited similar intracellular transport properties and all were expressed abundantly on the cell surface. Quantitative and qualitative cell fusion assays indicated that all of the F protein COOH-terminal truncation mutants mediated lipid mixing with similar kinetics and efficiency as that of wild-type F protein. However, the cytoplasmic content mixing activity decreased in parallel with the extent of the deletion in the F protein COOH-terminal truncation mutants. These data indicate that it is possible to separate the presumptive early step in the fusion reaction, hemifusion, and the final stage of fusion, content mixing, and that the presence of the F protein COOH-terminal region is important for the final steps of fusion.
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页码:73 / 84
页数:12
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共 75 条
[1]  
AHKONG QF, 1987, J CELL SCI, V88, P389
[2]   INDIVIDUAL ROLES OF N-LINKED OLIGOSACCHARIDE CHAINS IN INTRACELLULAR-TRANSPORT OF THE PARAMYXOVIRUS SV5 FUSION PROTEIN [J].
BAGAI, S ;
LAMB, RA .
VIROLOGY, 1995, 209 (01) :250-256
[3]   HEMAGGLUTININ-NEURAMINIDASE ENHANCES-F PROTEIN-MEDIATED MEMBRANE-FUSION OF RECONSTITUTED SENDAI VIRUS ENVELOPES WITH CELLS [J].
BAGAI, S ;
PURI, A ;
BLUMENTHAL, R ;
SARKAR, DP .
JOURNAL OF VIROLOGY, 1993, 67 (06) :3312-3318
[4]   QUANTITATIVE MEASUREMENT OF PARAMYXOVIRUS FUSION - DIFFERENCES IN REQUIREMENTS OF GLYCOPROTEINS BETWEEN SIMIAN-VIRUS-5 AND HUMAN PARAINFLUENZA-VIRUS-3 OR NEWCASTLE-DISEASE VIRUS [J].
BAGAI, S ;
LAMB, RA .
JOURNAL OF VIROLOGY, 1995, 69 (11) :6712-6719
[5]  
Brasseur R, 1988, Virus Genes, V1, P325
[6]   STRUCTURE OF INFLUENZA HEMAGGLUTININ AT THE PH OF MEMBRANE-FUSION [J].
BULLOUGH, PA ;
HUGHSON, FM ;
SKEHEL, JJ ;
WILEY, DC .
NATURE, 1994, 371 (6492) :37-43
[7]   A SPRING-LOADED MECHANISM FOR THE CONFORMATIONAL CHANGE OF INFLUENZA HEMAGGLUTININ [J].
CARR, CM ;
KIM, PS .
CELL, 1993, 73 (04) :823-832
[8]   HEPTAD REPEAT SEQUENCES ARE LOCATED ADJACENT TO HYDROPHOBIC REGIONS IN SEVERAL TYPES OF VIRUS FUSION GLYCOPROTEINS [J].
CHAMBERS, P ;
PRINGLE, CR ;
EASTON, AJ .
JOURNAL OF GENERAL VIROLOGY, 1990, 71 :3075-3080
[9]   THE HEMIFUSION INTERMEDIATE AND ITS CONVERSION TO COMPLETE FUSION - REGULATION BY MEMBRANE-COMPOSITION [J].
CHERNOMORDIK, L ;
CHANTURIYA, A ;
GREEN, J ;
ZIMMERBERG, J .
BIOPHYSICAL JOURNAL, 1995, 69 (03) :922-929
[10]   SITE-DIRECTED MUTAGENESIS OF VIRTUALLY ANY PLASMID BY ELIMINATING A UNIQUE SITE [J].
DENG, WP ;
NICKOLOFF, JA .
ANALYTICAL BIOCHEMISTRY, 1992, 200 (01) :81-88