Roles of Asp179 and Glu270 in ADP-Ribosylation of Actin by Clostridium perfringens Iota Toxin

被引：12
作者：
Belyy, Alexander [1 ,2 ]
Tabakova, Irina [1 ]
Lang, Alexander E. [3 ]
Jank, Thomas [3 ]
Belyi, Yury [1 ,4 ]
Aktories, Klaus [3 ,4 ,5 ]
机构：
[1] Gamaleya Res Inst, Moscow, Russia
[2] Moscow MV Lomonosov State Univ, Fac Biol, Moscow, Russia
[3] Univ Freiburg, Inst Expt & Clin Pharmacol & Toxicol, D-79106 Freiburg, Germany
[4] Univ Freiburg, Freiburg Inst Adv Studies FRIAS, D-79106 Freiburg, Germany
[5] Univ Freiburg, Ctr Biol Signalling Studies BIOSS, D-79106 Freiburg, Germany
来源：
PLOS ONE | 2015年 / 10卷 / 12期
关键词：
YEAST ACTIN; SACCHAROMYCES-CEREVISIAE; ENZYMATIC COMPONENT; SHUTTLE VECTORS; CELLULAR UPTAKE; BINDING; RECOGNITION; RESIDUES; MUTATION; MUTANTS;
D O I：
10.1371/journal.pone.0145708
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
Clostridium perfringens iota toxin is a binary toxin composed of the enzymatically active component Ia and receptor binding component Ib. Ia is an ADP-ribosyltransferase, which modifies Arg177 of actin. The previously determined crystal structure of the actin-Ia complex suggested involvement of Asp179 of actin in the ADP-ribosylation reaction. To gain more insights into the structural requirements of actin to serve as a substrate for toxin-catalyzed ADP-ribosylation, we engineered Saccharomyces cerevisiae strains, in which wild type actin was replaced by actin variants with substitutions in residues located on the Ia-actin interface. Expression of the actin mutant Arg177Lys resulted in complete resistance towards Ia. Actin mutation of Asp179 did not change Ia-induced ADP-ribosylation and growth inhibition of S. cerevisiae. By contrast, substitution of Glu270 of actin inhibited the toxic action of Ia and the ADP-ribosylation of actin. In vitro transcribed/translated human a-actin confirmed the crucial role of Glu270 in ADP-ribosylation of actin by Ia.
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页数：15
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