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PMID:9111062

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Citation

Zhang, S and Grosse, F (1997) Domain structure of human nuclear DNA helicase II (RNA helicase A). J. Biol. Chem. 272:11487-94

Abstract

Full-length human nuclear DNA helicase II (NDH II) was cloned and overexpressed in a baculovirus-derived expression system. Recombinant NDH II unwound both DNA and RNA. Limited tryptic digestion produced active helicases with molecular masses of 130 and 100 kDa. The 130-kDa helicase missed a glycine-rich domain (RGG-box) at the carboxyl terminus, while the 100-kDa form missed both its double-stranded RNA binding domains (dsRBDs) at the amino terminus and its RGG-box. Hence, the dsRBDs and the RGG-box were dispensable for unwinding. On the other hand, the isolated DEXH core alone could neither hydrolyze ATP nor unwind nucleic acids. These enzymatic activities were not regained by fusing a complete COOH or NH2 terminus to the helicase core. Hence, an active helicase required part of the NH2 terminus, the DEXH core, and a C-terminal extension of the core. Both dsRBDs and the RGG-box were bacterially expressed as glutathione S-transferase fusion proteins. The two dsRBDs had a strong affinity to double-stranded RNA and cooperated upon RNA binding, while the RGG-box bound preferentially to single-stranded DNA. A model is suggested in which the flanking domains influence and regulate the unwinding properties of NDH II.

Links

PubMed

Keywords

Adenosine Triphosphatases/chemistry; Adenosine Triphosphatases/genetics; Adenosine Triphosphatases/metabolism; Adenosine Triphosphate/metabolism; Amino Acid Sequence; Animals; Baculoviridae/genetics; Cattle; Cell Nucleus/enzymology; Cloning, Molecular; DNA Helicases/chemistry; DNA Helicases/genetics; DNA Helicases/metabolism; DNA, Single-Stranded/metabolism; DNA-Binding Proteins/metabolism; Humans; Models, Molecular; Molecular Sequence Data; Nucleic Acid Conformation; Peptide Fragments/metabolism; Protein Conformation; RNA Helicases; RNA Nucleotidyltransferases/chemistry; RNA Nucleotidyltransferases/genetics; RNA Nucleotidyltransferases/metabolism; RNA, Double-Stranded/metabolism; RNA-Binding Proteins/metabolism; Recombinant Proteins/metabolism; Sequence Analysis, DNA; T-Lymphocytes/enzymology

Significance

Annotations

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

HUMAN:DHX9

located_in

GO:0005634: nucleus

ECO:0000304: author statement supported by traceable reference used in manual assertion

C

Seeded From UniProt

complete

HUMAN:DHX9

enables

GO:0003678: DNA helicase activity

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

HUMAN:DHX9

enables

GO:0003724: RNA helicase activity

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

HUMAN:DHX9

part_of

GO:0005634: nucleus

ECO:0000304: author statement supported by traceable reference used in manual assertion

C

Seeded From UniProt

complete

HUMAN:DHX9

enables

GO:0004004: ATP-dependent RNA helicase activity

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

HUMAN:DHX9

enables

GO:0004003: ATP-dependent DNA helicase activity

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

HUMAN:DHX9

involved_in

GO:0032508: DNA duplex unwinding

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DHX9

involved_in

GO:0010501: RNA secondary structure unwinding

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DHX9

enables

GO:0003690: double-stranded DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:DHX9

enables

GO:0003697: single-stranded DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:DHX9

enables

GO:0003727: single-stranded RNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:DHX9

enables

GO:0003725: double-stranded RNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

Notes

See also

References

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