Association of human DNA helicase RecQ5β with RNA polymerase II and its possible role in transcription

被引:45
作者
Izumikawa, Keiichi [1 ]
Yanagida, Mitsuaki [1 ]
Hayano, Toshiya [1 ]
Tachikawa, Hiroyuki [1 ]
Komatsu, Wataru [1 ]
Shimamoto, Akira [2 ]
Futami, Kazunobu [2 ]
Furuichi, Yasuhiro [2 ]
Shinkawa, Takashi [3 ]
Yamauchi, Yoshio [3 ]
Isobe, Toshiaki [3 ,4 ]
Takahashi, Nobuhiro [1 ,4 ]
机构
[1] Tokyo Univ Agr & Technol, Dept Bioengn, United Grad Sch Agr, Fuchu, Tokyo 1838509, Japan
[2] GeneCare Res Inst, Kanagawa 2470063, Japan
[3] Tokyo Metropolitan Univ, Grad Sch Sci, Biochem Lab, Tokyo 1920397, Japan
[4] CREST, Japan Sci & Technol Agcy, Chuo Ku, Tokyo 1030027, Japan
关键词
functional proteomics; gene regulation; protein-protein interaction; RecQ5; beta; RNA polymerase II (RNAP II); transcription;
D O I
10.1042/BJ20071392
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
Although RecQ5 beta is a ssDNA (single-stranded DNA)-stimulated ATPase and an ATP-dependent DNA helicase with strand-annealing activities, its cellular function remains to be explored. In the present paper, we used immunopurification and MS-based analyses to show that human DNA helicase RecQ5 beta is associated with at least four RNAP II (RNA polymerase II) subunits. RecQ5 beta was also present in complexes immunoprecipitated using three different antibodies against the large subunit of RNAP II, or in complexes immunoprecipitated using an anti-FLAG antibody against either FLAG-RNAP II 33 kDa subunit or FLAG-Pin 1. Different regions of the non-helicase domain of the RecQ5 beta molecule were associated with hypophosphorylated and hyperphosphorylated forms of the RNAP II large subunit independently of DNA and RNA. RecQ5 beta was also found in nuclear chromatin fractions and associated with the coding regions of the LDL (low-density lipoprotem) receptor and beta-actin genes. Knockdown of the RecQ5 beta transcript increased the transcription of those genes. The results of the present study suggest that RecQ5 beta has suppressive roles in events associated with RNAP II-dependent transcription.
引用
收藏
页码:505 / 516
页数:12
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