Plasmodium falciparum RuvB1 is an active DNA helicase and translocates in the 5¡ä¨C3¡ä direction

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• 作者：Moaz Ahmad ; Renu Tuteja ; ^{renu@icgeb.res.in} ; ^{renututeja@gmail.com}
• 关键词：AAA  ; + ; ATPases associated with various cellular activities ; DAB ; 3 ; 3&prime ; -diaminobenzidine ; DAPI ; 40 ; 60-di-amidino-2-phenylindole dihydrochloride ; HRP ; horseradish peroxidase ; IPTG ; isopropyl-¦Â-d-thiogalactopyranoside ; PBS ; phosphate buffered sali
• 刊名：Gene
• 出版年：2013
• 出版时间：15 February, 2013
• 年：2013
• 卷：515
• 期：1
• 页码：99-109
• 全文大小：1201 K

文摘

RuvB family of protein contains two similar kinds of proteins i.e. RuvB1 and RuvB2 from yeast to human. These proteins belong to the AAA + class of proteins and are critical components of several multiprotein complexes involved in diverse cellular activities. There are two RuvB proteins annotated in the Plasmodium database but the identification of the third protein recently by our lab has raised the question why Plasmodium falciparum contains three RuvB proteins instead of two. Hence the biochemical characterizations of these proteins have become essential to understand the role of these proteins in the malaria parasite. Recently we have reported the characterization of the recombinant PfRuvB3, which contains ATPase activity but lacks DNA helicase activity. In the present study we report the phylogenetic analysis and detailed biochemical characterization of one of the other RuvB homologue RuvB1 from P. falciparum. PfRuvB1 shows considerable homology with human as well as yeast RuvB1 and contains Walker motif A and Walker motif B. The activity analysis of this protein revealed that PfRuvB1 is an ATPase and this activity increased significantly in the presence of ss-DNA. PfRuvB1 also contains DNA helicase activity and translocates preferentially in 5¡ä to 3¡ä direction. In vivo investigation of PfRuvB1 revealed that it is constitutively expressed during all the stages of intraerythrocytic cycle of P. falciparum and localizes mainly to the nucleus. These studies will make important contribution in understanding the role of RuvB protein in P. falciparum.