The V-ATPase subunit C binds to polymeric F-actin as well as to monomeric G-actin and induces cross-linking of actin filaments

被引:79
作者
Vitavska, O [1 ]
Merzendorfer, H [1 ]
Wieczorek, H [1 ]
机构
[1] Univ Osnabruck, Dept Biol Chem, Div Anim Physiol, D-49069 Osnabruck, Germany
关键词
D O I
10.1074/jbc.M406797200
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
Previously, we have shown that the V-ATPase holoenzyme as well as the V-1 complex isolated from the midgut of the tobacco hornworm ( Manduca sexta) exhibits the ability of binding to actin filaments via the V-1 subunits B and C (Vitavska, O., Wieczorek, H., and Merzendorfer, H. (2003) J. Biol. Chem. 278, 18499 - 18505). Since the recombinant subunit C not only enhances actin binding of the V-1 complex but also can bind separately to F-actin, we analyzed the interaction of recombinant subunit C with actin. We demonstrate that it binds not only to F-actin but also to monomeric G-actin. With dissociation constants of similar to 50 nM, the interaction exhibits a high affinity, and no difference could be observed between binding to ATP-G-actin or ADP-G-actin, respectively. Unlike other proteins such as members of the ADF/cofilin family, which also bind to G- as well as to F-actin, subunit C does not destabilize actin filaments. On the contrary, under conditions where the disassembly of F-actin into G- actin usually occurred, subunit C stabilized F-actin. In addition, it increased the initial rate of actin polymerization in a concentration-dependent manner and was shown to cross-link actin filaments to bundles of varying thickness. Apparently bundling is enabled by the existence of at least two actin-binding sites present in the N- and in the C-terminal halves of subunits C, respectively. Since subunit C has the possibility to dimerize or even to oligomerize, spacing between actin filaments could be variable in size.
引用
收藏
页码:1070 / 1076
页数:7
相关论文
共 38 条
[1]   Structural analysis of the stalk subunit Vma5p of the yeast V-ATPase in solution [J].
Armbrüster, A ;
Svergun, DI ;
Coskun, U ;
Juliano, S ;
Bailer, SM ;
Grüber, G .
FEBS LETTERS, 2004, 570 (1-3) :119-125
[2]   In vivo functions of actin-binding proteins [J].
Ayscough, KR .
CURRENT OPINION IN CELL BIOLOGY, 1998, 10 (01) :102-111
[3]   TECHNIQUES FOR REARING LABORATORY COLONIES OF TOBACCO HORNWORMS AND PINK BOLLWORMS LEPIDOPTERA-SPHINGIDAE-GELECHIIDAE [J].
BELL, RA ;
JOACHIM, FG .
ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA, 1976, 69 (02) :365-373
[4]  
BONFANTI P, 1992, EUR J CELL BIOL, V57, P298
[5]   Macromolecular crowding as a cell volume sensor [J].
Burg, MB .
CELLULAR PHYSIOLOGY AND BIOCHEMISTRY, 2000, 10 (5-6) :251-256
[6]   Actin depolymerizing factor (ADF/cofilin) enhances the rate of filament turnover: Implication in actin-based motility [J].
Carlier, MF ;
Laurent, V ;
Santolini, J ;
Melki, R ;
Didry, D ;
Xia, GX ;
Hong, Y ;
Chua, NH ;
Pantaloni, D .
JOURNAL OF CELL BIOLOGY, 1997, 136 (06) :1307-1322
[7]   Vacuolar H+-ATPase binding to microfilaments -: Regulation in response to phosphatidylinositol 3-kinase activity and detailed characterization of the actin-binding site in subunit B [J].
Chen, SH ;
Bubb, MR ;
Yarmola, EG ;
Zuo, J ;
Jiang, J ;
Lee, BS ;
Lu, M ;
Gluck, SL ;
Hurst, IR ;
Holliday, LS .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (09) :7988-7998
[8]  
COOPER JA, 1982, METHOD ENZYMOL, V85, P182
[9]  
DETMERS P, 1981, J BIOL CHEM, V256, P99
[10]   Phosphoinositide binding inhibits α-actinin bundling activity [J].
Fraley, TS ;
Tran, TC ;
Corgan, AM ;
Nash, CA ;
Hao, J ;
Critchley, DR ;
Greenwood, JA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (26) :24039-24045