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PMID:15123714

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Citation

Rundle, DR, Gorbsky, G and Tsiokas, L (2004) PKD2 interacts and co-localizes with mDia1 to mitotic spindles of dividing cells: role of mDia1 IN PKD2 localization to mitotic spindles. J. Biol. Chem. 279:29728-39

Abstract

Mutations in pkd2 result in the type 2 form of autosomal dominant polycystic kidney disease, which accounts for approximately 15% of all cases of the disease. PKD2, the protein product of pkd2, belongs to the transient receptor potential superfamily of cation channels, and it can function as a mechanosensitive channel in the primary cilium of kidney cells, an intracellular Ca(2+) release channel in the endoplasmic reticulum, and/or a nonselective cation channel in the plasma membrane. We have identified mDia1/Drf1 (mammalian Diaphanous or Diaphanous-related formin 1 protein) as a PKD2-interacting protein by yeast two-hybrid screen. mDia1 is a member of the RhoA GTPase-binding formin homology protein family that participates in cytoskeletal organization, cytokinesis, and signal transduction. We show that mDia1 and PKD2 interact in native and in transfected cells, and binding is mediated by the cytoplasmic C terminus of PKD2 binding to the mDia1 N terminus. The interaction is more prevalent in dividing cells in which endogenous PKD2 and mDia1 co-localize to the mitotic spindles. RNA interference experiments reveal that endogenous mDia1 knockdown in HeLa cells results in the loss of PKD2 from mitotic spindles and alters intracellular Ca(2+) release. Our results suggest that mDia1 facilitates the movement of PKD2 to a centralized position during cell division and has a positive effect on intracellular Ca(2+) release during mitosis. This may be important to ensure equal segregation of PKD2 to the daughter cell to maintain a necessary level of channel activity. Alternatively, PKD2 channel activity may be important in the cell division process or in cell fate decisions after division.

Links

PubMed Online version:10.1074/jbc.M400544200

Keywords

Animals; Calcium/metabolism; Calcium Channels/genetics; Calcium Channels/metabolism; Cell Division/physiology; HeLa Cells; Humans; Immunohistochemistry; Male; Membrane Proteins/genetics; Membrane Proteins/metabolism; Microtubules/metabolism; Mitotic Spindle Apparatus/metabolism; Protein Structure, Tertiary; Recombinant Fusion Proteins/metabolism; TRPP Cation Channels; Tubulin/genetics; Tubulin/metabolism; Two-Hybrid System Techniques

Significance

Annotations

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

PIG:F1RMT5

located_in

GO:0015630: microtubule cytoskeleton

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

PIG:F1RMT5

colocalizes_with

GO:0005876: spindle microtubule

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

PIG:F2VYA1

located_in

GO:0097730: non-motile cilium

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

PIG:F2VYA1

located_in

GO:0015630: microtubule cytoskeleton

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

PIG:F2VYA1

colocalizes_with

GO:0005876: spindle microtubule

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

PIG:F2VYA1

located_in

GO:0072686: mitotic spindle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:DIAP1

enables

GO:0044325: transmembrane transporter binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q13563

F

Seeded From UniProt

complete

MOUSE:PKD2

located_in

GO:0072686: mitotic spindle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:DIAP1

located_in

GO:0072686: mitotic spindle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:DIAP1

enables

GO:0044325: transmembrane transporter binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q13563

F

Seeded From UniProt

complete

HUMAN:DIAP1

involved_in

GO:0051279: regulation of release of sequestered calcium ion into cytosol

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DIAP1

involved_in

GO:0071420: cellular response to histamine

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:DIAP1

involved_in

GO:0035372: protein localization to microtubule

ECO:0000315: mutant phenotype evidence used in manual assertion

P

  • has_input:(UniProtKB:Q13563)

Seeded From UniProt

complete

HUMAN:PKD2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O60610

F

Seeded From UniProt

complete

HUMAN:PKD2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O08808

F

Seeded From UniProt

complete

HUMAN:PKD2

located_in

GO:0005783: endoplasmic reticulum

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:PKD2

located_in

GO:0072686: mitotic spindle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:PKD2

part_of

GO:0072686: mitotic spindle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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