Biochemical characterization of Rous sarcoma virus MA protein interaction with membranes

被引:57
作者
Dalton, AK
Murray, PS
Murray, D
Vogt, VM
机构
[1] Cornell Univ, Dept Mol Biol & Genet, Ithaca, NY 14853 USA
[2] Weill Corell Med Coll, Dept Microbiol & Immunol, New York, NY 10021 USA
[3] Weill Corell Med Coll, Inst Computat Biomed, New York, NY 10021 USA
关键词
D O I
10.1128/JVI.79.10.6227-6238.2005
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The MA domain of retroviral Gag proteins mediates association with the host cell membrane during assembly. The biochemical nature of this interaction is not well understood. We have used an in vitro flotation assay to directly measure Rous sarcoma virus (RSV) MA-membrane interaction in the absence of host cell factors. The association of purified MA and MA-containing proteins with liposomes of defined composition was electrostatic in nature and depended upon the presence of a biologically relevant concentration of negatively charged lipids. A mutant MA protein known to be unable to promote Gag membrane association and budding in vivo failed to bind to liposomes. These results were supported by computational modeling. The intrinsic affinity of RSV MA for negatively charged membranes appears insufficient to promote efficient plasma membrane binding during assembly. However, an artificially dimerized form of MA bound to liposomes by at least an order of magnitude more tightly than monomeric MA. This result suggests that the clustering of MA domains, via Gag-Gag interactions during virus assembly, drives membrane association in vivo.
引用
收藏
页码:6227 / 6238
页数:12
相关论文
共 98 条
[1]  
Alexandrov N N, 1996, Pac Symp Biocomput, P53
[2]   LIPID-COMPOSITION AND FLUIDITY OF THE HUMAN-IMMUNODEFICIENCY-VIRUS ENVELOPE AND HOST-CELL PLASMA-MEMBRANES [J].
ALOIA, RC ;
TIAN, HR ;
JENSEN, FC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (11) :5181-5185
[3]  
Baker Nathan A, 2003, Methods Biochem Anal, V44, P427
[4]   FUNCTIONAL CHIMERAS OF THE ROUS-SARCOMA VIRUS AND HUMAN-IMMUNODEFICIENCY-VIRUS GAG PROTEINS [J].
BENNETT, RP ;
NELLE, TD ;
WILLS, JW .
JOURNAL OF VIROLOGY, 1993, 67 (11) :6487-6498
[5]   Electrostatic binding of proteins to membranes. Theoretical predictions and experimental results with charybdotoxin and phospholipid vesicles [J].
BenTal, N ;
Honig, B ;
Miller, C ;
McLaughlin, S .
BIOPHYSICAL JOURNAL, 1997, 73 (04) :1717-1727
[6]   Binding of small basic peptides to membranes containing acidic lipids: Theoretical models and experimental results [J].
BenTal, N ;
Honig, B ;
Peitzsch, RM ;
Denisov, G ;
McLaughlin, S .
BIOPHYSICAL JOURNAL, 1996, 71 (02) :561-575
[7]   CHARMM - A PROGRAM FOR MACROMOLECULAR ENERGY, MINIMIZATION, AND DYNAMICS CALCULATIONS [J].
BROOKS, BR ;
BRUCCOLERI, RE ;
OLAFSON, BD ;
STATES, DJ ;
SWAMINATHAN, S ;
KARPLUS, M .
JOURNAL OF COMPUTATIONAL CHEMISTRY, 1983, 4 (02) :187-217
[8]   MYRISTOYLATION-DEPENDENT REPLICATION AND ASSEMBLY OF HUMAN IMMUNODEFICIENCY VIRUS-1 [J].
BRYANT, M ;
RATNER, L .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (02) :523-527
[9]   MEMBRANE-BINDING OF MYRISTYLATED PEPTIDES CORRESPONDING TO THE NH2 TERMINUS OF SRC [J].
BUSER, CA ;
SIGAL, CT ;
RESH, MD ;
MCLAUGHLIN, S .
BIOCHEMISTRY, 1994, 33 (44) :13093-13101
[10]   HMMSTR: a hidden Markov model for local sequence-structure correlations in proteins [J].
Bystroff, C ;
Thorsson, V ;
Baker, D .
JOURNAL OF MOLECULAR BIOLOGY, 2000, 301 (01) :173-190