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PMID:9817760

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Citation

Nakamura, M, Masuda, H, Horii, J, Kuma, K, Yokoyama, N, Ohba, T, Nishitani, H, Miyata, T, Tanaka, M and Nishimoto, T (1998) When overexpressed, a novel centrosomal protein, RanBPM, causes ectopic microtubule nucleation similar to gamma-tubulin. J. Cell Biol. 143:1041-52

Abstract

A novel human protein with a molecular mass of 55 kD, designated RanBPM, was isolated with the two-hybrid method using Ran as a bait. Mouse and hamster RanBPM possessed a polypeptide identical to the human one. Furthermore, Saccharomyces cerevisiae was found to have a gene, YGL227w, the COOH-terminal half of which is 30% identical to RanBPM. Anti-RanBPM antibodies revealed that RanBPM was localized within the centrosome throughout the cell cycle. Overexpression of RanBPM produced multiple spots which were colocalized with gamma-tubulin and acted as ectopic microtubule nucleation sites, resulting in a reorganization of microtubule network. RanBPM cosedimented with the centrosomal fractions by sucrose- density gradient centrifugation. The formation of microtubule asters was inhibited not only by anti- RanBPM antibodies, but also by nonhydrolyzable GTP-Ran. Indeed, RanBPM specifically interacted with GTP-Ran in two-hybrid assay. The central part of asters stained by anti-RanBPM antibodies or by the mAb to gamma-tubulin was faded by the addition of GTPgammaS-Ran, but not by the addition of anti-RanBPM anti- bodies. These results provide evidence that the Ran-binding protein, RanBPM, is involved in microtubule nucleation, thereby suggesting that Ran regulates the centrosome through RanBPM.

Links

PubMed PMC2132962

Keywords

Adaptor Proteins, Signal Transducing; Amino Acid Sequence; Animals; CHO Cells; COS Cells; Centrosome/chemistry; Centrosome/drug effects; Cricetinae; Cytoskeletal Proteins; DNA, Complementary; Evolution, Molecular; GTP-Binding Proteins/analysis; GTP-Binding Proteins/chemistry; GTP-Binding Proteins/genetics; Gene Expression/physiology; Guanosine 5'-O-(3-Thiotriphosphate)/pharmacology; HeLa Cells; Humans; Immunoblotting; Microtubules/chemistry; Molecular Sequence Data; Nuclear Proteins/analysis; Nuclear Proteins/chemistry; Nuclear Proteins/genetics; Tubulin/chemistry; Tubulin/genetics; ran GTP-Binding Protein

Significance

Annotations

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

HUMAN:RANB9

enables

GO:0031267: small GTPase binding

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

F

Seeded From UniProt

complete

HUMAN:RANB9

part_of

GO:0005875: microtubule associated complex

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

C

Seeded From UniProt

complete

HUMAN:RANB9

involved_in

GO:0007020: microtubule nucleation

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

P

Seeded From UniProt

complete

HUMAN:RANB9

enables

GO:0008536: Ran GTPase binding

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

F

Seeded From UniProt

complete

HUMAN:RANB9

involved_in

GO:0065003: protein-containing complex assembly

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

P

Seeded From UniProt

complete


See also

References

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