SITE-DIRECTED MUTAGENESIS OF THE TICK-BORNE ENCEPHALITIS-VIRUS NS3 GENE REVEALS THE PUTATIVE SERINE-PROTEASE DOMAIN OF THE NS3 PROTEIN

被引:12
作者
PUGACHEV, KV
NOMOKONOVA, NY
DOBRIKOVA, EY
WOLF, YI
机构
[1] RUSSIAN ACAD SCI,INST BIOORGAN CHEM,SIBERIAN DIV,NOVOSIBIRSK 630090,RUSSIA
[2] RUSSIAN ACAD MED SCI,INST THERAPY,SIBERIAN DIV,NOVOSIBIRSK 630003,RUSSIA
[3] RUSSIAN ACAD SCI,INST CYTOL & GENET,SIBERIAN DIV,NOVOSIBIRSK 630090,RUSSIA
关键词
FLAVIVIRUS; NONSTRUCTURAL PROTEIN; SERINE PROTEASE; CLEAVAGE;
D O I
10.1016/0014-5793(93)80977-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Several mutations were introduced into the putative serine protease domain of the tick-borne encephalitis virus NS3 protein and into a possible internal cleavage site within the protein. The influence of these mutations on proteolytic activity of NS3 protein and NS3' protein formation was tested in vitro. It was found that NS3' formation was not dependent on the activity of the NS3 N-terminal serine protease. Mutations affecting the Ser-138 residue of the NS3 protein prohibited cleavage between NS2B and NS3 proteins when the NS2B-NS3 part of the viral genome was expressed in vitro, suggesting the key role of Ser-138 in viral serine protease functioning.
引用
收藏
页码:115 / 118
页数:4
相关论文
共 16 条
[1]   DETECTION OF A TRYPSIN-LIKE SERINE PROTEASE DOMAIN IN FLAVIVIRUSES AND PESTIVIRUSES [J].
BAZAN, JF ;
FLETTERICK, RJ .
VIROLOGY, 1989, 171 (02) :637-639
[2]   EVIDENCE THAT THE N-TERMINAL DOMAIN OF NONSTRUCTURAL PROTEIN NS3 FROM YELLOW-FEVER VIRUS IS A SERINE PROTEASE RESPONSIBLE FOR SITE-SPECIFIC CLEAVAGES IN THE VIRAL POLYPROTEIN [J].
CHAMBERS, TJ ;
WEIR, RC ;
GRAKOUI, A ;
MCCOURT, DW ;
BAZAN, JF ;
FLETTERICK, RJ ;
RICE, CM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (22) :8898-8902
[3]  
DAYHOFF MO, 1983, METHOD ENZYMOL, V91, P524
[4]   PROPER PROCESSING OF DENGUE VIRUS NONSTRUCTURAL GLYCOPROTEIN-NS1 REQUIRES THE N-TERMINAL HYDROPHOBIC SIGNAL SEQUENCE AND THE DOWNSTREAM NONSTRUCTURAL PROTEIN-NS2A [J].
FALGOUT, B ;
CHANOCK, R ;
LAI, CJ .
JOURNAL OF VIROLOGY, 1989, 63 (05) :1852-1860
[5]   BOTH NONSTRUCTURAL PROTEINS NS2B AND NS3 ARE REQUIRED FOR THE PROTEOLYTIC PROCESSING OF DENGUE VIRUS NONSTRUCTURAL PROTEINS [J].
FALGOUT, B ;
PETHEL, M ;
ZHANG, YM ;
LAI, CJ .
JOURNAL OF VIROLOGY, 1991, 65 (05) :2467-2475
[6]   N-TERMINAL DOMAINS OF PUTATIVE HELICASES OF FLAVIVIRUSES AND PESTIVIRUSES MAY BE SERINE PROTEASES [J].
GORBALENYA, AE ;
DONCHENKO, AP ;
KOONIN, EV ;
BLINOV, VM .
NUCLEIC ACIDS RESEARCH, 1989, 17 (10) :3889-3897
[7]   CLEAVAGE OF DENGUE VIRUS NS1-NS2A REQUIRES AN OCTAPEPTIDE SEQUENCE AT THE C-TERMINUS OF NS1 [J].
HORI, H ;
LAI, CJ .
JOURNAL OF VIROLOGY, 1990, 64 (09) :4573-4577
[8]   THE GAPPED DUPLEX DNA APPROACH TO OLIGONUCLEOTIDE-DIRECTED MUTATION CONSTRUCTION [J].
KRAMER, W ;
DRUTSA, V ;
JANSEN, HW ;
KRAMER, B ;
PFLUGFELDER, M ;
FRITZ, HJ .
NUCLEIC ACIDS RESEARCH, 1984, 12 (24) :9441-9456
[9]  
KUNKEL TA, 1987, METHOD ENZYMOL, V154, P367
[10]  
MAKAROVA K S, 1992, Molekulyarnaya Biologiya (Moscow), V26, P93