Characterization of the nucleic acid binding activity of inner core protein VP6 of African horse sickness virus

被引:9
作者
de Waal, PJ [1 ]
Huismans, H [1 ]
机构
[1] Univ Pretoria, Dept Genet, ZA-0002 Pretoria, South Africa
关键词
D O I
10.1007/s00705-005-0547-4
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Minor structural protein VP6 is the putative helicase of African horse sickness virus (AHSV), of the genus Orbivirus in the Reoviridae family. We investigated how the protein interacts with double-stranded (ds) RNA and other nucleic acids. Binding was assayed using an electrophoretic migration retardation assay and a nucleic acid overlay protein blot assay. VP6 bound double and single stranded RNA and DNA in a NaCl concentration sensitive reaction. Of six truncated VP6 peptides investigated, two partially overlapping peptides were found to bind dsRNA at pH 7.0, while other peptides with the same overlap did not. The distinction between the peptides appeared to be the pI which ranged from more than 8.0 to just above 6.0. Changing the pH of the binding buffer modified the binding activity. Regardless of assay conditions, only peptides with a specific region of amino acids in common, showed evidence of binding activity. No sequence homology was identified with other binding domains, however, the presence of charged amino acids are assumed to be important for binding activity. The results suggested dsRNA binding in the blot assay was strongly affected by the net charge on the peptide.
引用
收藏
页码:2037 / 2050
页数:14
相关论文
共 25 条
[1]   CHARACTERIZATION AND CLONING OF THE AFRICAN HORSESICKNESS VIRUS GENOME [J].
BREMER, CW ;
HUISMANS, H ;
VANDIJK, AA .
JOURNAL OF GENERAL VIROLOGY, 1990, 71 :793-799
[2]  
Calisher CH, 1998, ARCH VIROL, P3
[3]   The RNA helicase CI from plum pox potyvirus has two regions involved in binding to RNA [J].
Fernandez, A ;
Garcia, JA .
FEBS LETTERS, 1996, 388 (2-3) :206-210
[4]   COMPLETION OF THE SEQUENCE OF BLUETONGUE VIRUS SEROTYPE-10 BY THE CHARACTERIZATION OF A STRUCTURAL PROTEIN, VP6, AND A NON-STRUCTURAL PROTEIN, NS2 [J].
FUKUSHO, A ;
YU, Y ;
YAMAGUCHI, S ;
ROY, P .
JOURNAL OF GENERAL VIROLOGY, 1989, 70 :1677-1689
[5]   The highly ordered double-stranded RNA genome of bluetongue virus revealed by crystallography [J].
Gouet, P ;
Diprose, JM ;
Grimes, JM ;
Malby, R ;
Burroughs, JN ;
Zientara, S ;
Stuart, DI ;
Mertens, PPC .
CELL, 1999, 97 (04) :481-490
[6]   The atomic structure of the bluetongue virus core [J].
Grimes, JM ;
Burroughs, JN ;
Gouet, P ;
Diprose, JM ;
Malby, R ;
Ziéntara, S ;
Mertens, PPC ;
Stuart, DI .
NATURE, 1998, 395 (6701) :470-478
[7]   Expression and functional characterization of bluetongue virus VP2 protein: Role in cell entry [J].
Hassan, SS ;
Roy, P .
JOURNAL OF VIROLOGY, 1999, 73 (12) :9832-9842
[8]  
HASSAN Y, 2001, P RSTGC 2001 INT C, V5, P75
[9]   MAPPING AND CHARACTERIZATION OF ANTIGENIC EPITOPES AND THE NUCLEIC ACID-BINDING DOMAINS OF THE VP6 PROTEIN OF BLUETONGUE VIRUSES [J].
HAYAMA, E ;
LI, JKK .
JOURNAL OF VIROLOGY, 1994, 68 (06) :3604-3611
[10]   AFRICAN HORSE SICKNESS [J].
HOUSE, JA .
VETERINARY CLINICS OF NORTH AMERICA-EQUINE PRACTICE, 1993, 9 (02) :355-364