Umbravirus-encoded movement protein induces tubule formation on the surface of protoplasts and binds RNA incompletely and non-cooperatively

被引:30
作者
Nurkiyanova, KM
Ryabov, EV
Kalinina, NO
Fan, YC
Andreev, I
Fitzgerald, AG
Palukaitis, P
Taliansky, M [1 ]
机构
[1] Scottish Crop Res Inst, Virol Unit, Dundee DD2 5DA, Scotland
[2] Univ Dundee, Dept Elect Engn & Phys, Dundee DD1 4NH, Scotland
[3] Moscow MV Lomonosov State Univ, Inst Nucl Physicochem Biol, Moscow 119899, Russia
关键词
D O I
10.1099/0022-1317-82-10-2579
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Various functions of the cell-to-cell movement protein (MP) of Groundnut rosette virus (GRV) were analysed. The GRV ORF4-encoded protein was shown by immunofluorescence microscopy to generate tubular structures that protrude from the surface of the protoplast. The protein encoded by ORF4 was assessed also for RNA-binding properties. This protein was tagged at its C terminus with six histidine residues, produced in Escherichia coli using an expression vector and purified by affinity chromatography. Gel retardation analysis demonstrated that, in contrast to many other viral MPs, including the 3a MP of Cucumber mosaic virus (CMV), the ORF4-encoded protein bound non-cooperatively to viral ssRNA and formed complexes of low protein: RNA ratios. Competition binding experiments showed that the ORF4-encoded protein bound to both ssRNA and ssDNA without sequence specificity, but did not bind to dsDNA. UV cross-linking and nitrocellulose membrane-retention assays confirmed that both the GRV and the CMV MPs formed complexes with ssRNA and that these complexes showed similar stability in NaCl. Probing the MP-RNA complexes by atomic force microscopy demonstrated that the ORF4-encoded protein bound RNA incompletely, leaving protein-free RNA segments of varying length, while the CMV 3a protein formed highly packed complexes. The significance of the two properties of limited RNA binding and tubule formation of the umbraviral MP is discussed.
引用
收藏
页码:2579 / 2588
页数:10
相关论文
共 59 条
[41]  
Powell J., 1985, Proceedings of the 2nd International Conference on Lasers in Manufacturing, P37
[42]   CONSTRUCTION OF FULL-LENGTH CDNA CLONES OF CUCUMBER MOSAIC-VIRUS RNA-1, RNA-2 AND RNA-3 - GENERATION OF INFECTIOUS RNA TRANSCRIPTS [J].
RIZZO, TM ;
PALUKAITIS, P .
MOLECULAR & GENERAL GENETICS, 1990, 222 (2-3) :249-256
[43]   Host-specific cell-to-cell and long-distance movements of cucumber mosaic virus are facilitated by the movement protein of groundnut rosette virus [J].
Ryabov, EV ;
Roberts, IM ;
Palukaitis, P ;
Taliansky, M .
VIROLOGY, 1999, 260 (01) :98-108
[44]   Intracellular location of two groundnut rosette umbravirus proteins delivered by PVX and TMV vectors [J].
Ryabov, EV ;
Oparka, KJ ;
Santa Cruz, S ;
Robinson, DJ ;
Taliansky, ME .
VIROLOGY, 1998, 242 (02) :303-313
[45]   A plant virus-encoded protein facilitates long-distance movement of heterologous viral RNA [J].
Ryabov, EV ;
Robinson, DJ ;
Taliansky, ME .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (04) :1212-1217
[46]   NUCLEIC ACID-BINDING PROPERTIES OF THE ALFALFA MOSAIC-VIRUS MOVEMENT PROTEIN PRODUCED IN YEAST [J].
SCHOUMACHER, F ;
ERNY, C ;
BERNA, A ;
GODEFROYCOLBURN, T ;
STUSSIGARAUD, C .
VIROLOGY, 1992, 188 (02) :896-899
[47]   Visualization of poly(A)-binding protein complex formation with poly(A) RNA using atomic force microscopy [J].
Smith, BL ;
Gallie, DR ;
Le, H ;
Hansma, PK .
JOURNAL OF STRUCTURAL BIOLOGY, 1997, 119 (02) :109-117
[48]   The nonstructural NSm protein of tomato spotted wilt virus induces tubular structures in plant and insect cells [J].
Storms, MMH ;
Kormelink, R ;
Peters, D ;
vanLent, JWM ;
Goldbach, RW .
VIROLOGY, 1995, 214 (02) :485-493
[49]   FUNCTIONAL-ANALYSIS OF DELETION MUTANTS OF CUCUMBER MOSAIC-VIRUS RNA3 USING AN INVITRO TRANSCRIPTION SYSTEM [J].
SUZUKI, M ;
KUWATA, S ;
KATAOKA, J ;
MASUTA, C ;
NITTA, N ;
TAKANAMI, Y .
VIROLOGY, 1991, 183 (01) :106-113
[50]   Groundnut rosette disease virus complex: Biology and molecular biology [J].
Taliansky, ME ;
Robinson, DJ ;
Murant, AF .
ADVANCES IN VIRUS RESEARCH, VOL 55, 2000, 55 :357-400