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PMID:23219504

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Citation

Ahmad, M and Tuteja, R (2013) Plasmodium falciparum RuvB1 is an active DNA helicase and translocates in the 5'-3' direction. Gene 515:99-109

Abstract

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' to 3' 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.

Links

PubMed Online version:10.1016/j.gene.2012.11.020

Keywords

Adenosine Triphosphatases/chemistry; Adenosine Triphosphatases/genetics; Adenosine Triphosphatases/metabolism; Amino Acid Sequence; Cloning, Molecular; DNA Helicases/chemistry; DNA Helicases/genetics; DNA Helicases/metabolism; Enzyme Activation; Gene Expression; Humans; Molecular Sequence Data; Phylogeny; Plasmodium falciparum/enzymology; Plasmodium falciparum/genetics; Protozoan Proteins/chemistry; Protozoan Proteins/genetics; Protozoan Proteins/metabolism; Recombinant Proteins/chemistry; Recombinant Proteins/genetics; Recombinant Proteins/metabolism; Sequence Alignment

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

PLAF7:Q8IAT2

enables

GO:0003678: DNA helicase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

PLAF7:Q8IAT2

enables

GO:0043139: 5'-3' DNA helicase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

PLAFA:J7HDV0

enables

GO:0043141: ATP-dependent 5'-3' DNA helicase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

PLAFA:J7HDV0

enables

GO:0008094: DNA-dependent ATPase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

PLAFA:J7HDV0

GO:0043141: ATP-dependent 5'-3' DNA helicase activity

ECO:0000314:

F

Fig. 5

complete
CACAO 8169

PLAFA:J7HDV0

GO:0008094: DNA-dependent ATPase activity

ECO:0000314:

F

Figure 4

complete
CACAO 8170

PLAFA:J7HDV0

GO:0005634: nucleus

ECO:0000314:

C

Fig. 6A

complete
CACAO 8185

PLAFA:J7HDV0

part_of

GO:0005634: nucleus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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