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PMID:11402034

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Citation

Yang, JP, Tang, H, Reddy, TR and Wong-Staal, F (2001) Mapping the functional domains of HAP95, a protein that binds RNA helicase A and activates the constitutive transport element of type D retroviruses. J. Biol. Chem. 276:30694-700

Abstract

The complex retroviruses such as human immunodeficiency virus, type 1, employ a virally encoded protein, Rev, to mediate the nuclear export of unspliced and partially spliced mRNA. In contrast, the simian type D retroviruses act through a cis-acting constitutive transport element (CTE) that presumably interacts directly with cellular export proteins. We first reported that RNA helicase A (RHA) is a shuttle protein that binds to functional CTE in vitro and in vivo. Recently, we isolated a novel protein, HAP95, that specifically binds to the nuclear transport domain of RHA and up-regulates CTE-mediated gene expression. Here, using truncation and deletion mutations, we mapped the domains of HAP95 that are important for RHA binding, transactivation of CTE, and nuclear cytoplasmic shuttling. We report evidence for a novel nuclear export signal in HAP95 and showed that the domains involved in RHA binding and nuclear localization are required for CTE activation. Finally, we showed that HAP95 synergizes significantly with RHA on CTE-mediated reporter gene expression and promotes nuclear export of unspliced mRNA in transfected cells. Taken together, these data support the proposal that HAP95 specifically facilitates CTE-mediated gene expression by directly binding to RHA.

Links

PubMed Online version:10.1074/jbc.M102809200

Keywords

Active Transport, Cell Nucleus; Amino Acid Sequence; Autoantigens/metabolism; Betaretrovirus/metabolism; Carrier Proteins/chemistry; Carrier Proteins/physiology; DEAD-box RNA Helicases; DNA-Binding Proteins; HeLa Cells; Humans; Intracellular Signaling Peptides and Proteins; Molecular Sequence Data; Neoplasm Proteins; Nuclear Proteins/chemistry; Nuclear Proteins/physiology; RNA/metabolism; RNA Helicases/metabolism

Significance

Annotations

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

HUMAN:DHX9

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9ULX6

F

Seeded From UniProt

complete

HUMAN:AKP8L

part_of

GO:1990904: ribonucleoprotein complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:AKP8L

located_in

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:AKP8L

located_in

GO:0005634: nucleus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:AKP8L

involved_in

GO:0010793: regulation of mRNA export from nucleus

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:AKP8L

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q08211

F

Seeded From UniProt

complete

HUMAN:DHX9

enables

GO:1990825: sequence-specific mRNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:DHX9

involved_in

GO:0046833: positive regulation of RNA export from nucleus

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DHX9

part_of

GO:1990904: ribonucleoprotein complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

Notes

See also

References

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