Screening for proteins interacting with ataxin-3, the gene product of SCA3/MJD.
- Author:
Lu SHEN
1
;
Bei-sha TANG
;
Jian-guang TANG
;
Hong JIANG
;
Cheng WANG
;
Hai-yan FANG
Author Information
1. Department of Neurology, Xiangya Hospital, China. shenlu2505@yahoo.com.cn
- Publication Type:Journal Article
- MeSH:
Ataxin-3;
Brain;
metabolism;
Gene Library;
Humans;
Nerve Tissue Proteins;
genetics;
metabolism;
Nuclear Proteins;
genetics;
metabolism;
Protein Interaction Mapping;
Repressor Proteins;
genetics;
metabolism;
Spinocerebellar Degenerations;
genetics;
metabolism;
Two-Hybrid System Techniques;
Yeasts;
genetics
- From:
Journal of Central South University(Medical Sciences)
2006;31(1):40-44
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To screen for proteins interacting with ataxin-3 by yeast two-hybrid system 3, and to discuss the function of ataxin-3 and pathogenesis of spinocerebellar ataxia type 3 and Machado-Joseph disease (SCA3/MJD).
METHODS:First we sub-cloned the full reading frame of both wild-type and mutant ataxin-3 into carrier pGBKT7 (ataxin-3-bait), and then screened human brain cDNA library with ataxin-3-bait.
RESULTS:We found five positive clones in 6.5 x 10(6) transformers. After sequencing, we knew all of them were novel ataxin-3 interacting proteins. Three were corresponded to the known sequences coding the known proteins, which were human Rho GDP dissociation inhibitor alpha, small ubiquitin-like modifier 1, and human neuronal amiloride-sensitive cation channel 2. Another two of the five were unknown.
CONCLUSION:Small ubiquitin-like modifier 1 probably interacted with ataxin-3, suggesting that the sumoylation probably participated in post-translation modifying of ataxin-3 and pathogenesis of SCA3/MJD.
