Tctex-1, a novel interaction partner of Rab3D, is required for osteoclastic bone resorption.
Vesicular transport along microtubules must be strictly regulated to sustain the unique structural and functional polarization of bone-resorbing osteoclasts. However, the molecular mechanisms bridging these vesicle-microtubule interactions remain largely obscure. Rab3D, a member of the Rab3 subfamily (Rab3A/B/C/D) of small exocytotic GTPases, represents a core component of the osteoclastic vesicle transport machinery. Here, we identify a new Rab3D-interacting partner, Tctex-1, a light chain of the cytoplasmic dynein microtubule motor complex, by a yeast two-hybrid screen. We demonstrate that Tctex-1 binds specifically to Rab3D in a GTP-dependent manner and co-occupies Rab3D-bearing vesicles in bone-resorbing osteoclasts. Furthermore, we provide evidence that Tctex-1 and Rab3D intimately associate with the dynein motor complex and microtubules in osteoclasts. Finally, targeted disruption of Tctex-1 by RNA interference significantly impairs bone resorption capacity and mislocalizes Rab3D vesicles in osteoclasts, attesting to the notion that components of the Rab3D-trafficking pathway contribute to the maintenance of osteoclastic resorptive function.
Centre for Orthopaedic Research, School of Surgery, University of Western Australia, Perth 6009, Western Australia, Australia.
Nathan J
NJ
Pavlos
Tak Sum
TS
Cheng
An
A
Qin
Pei Ying
PY
Ng
Hao-Tian
HT
Feng
Estabelle S M
ES
Ang
Amerigo
A
Carrello
Ching-Hwa
CH
Sung
Reinhard
R
Jahn
Ming-Hao
MH
Zheng
Jiake
J
Xu
D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport; D001862:Bone Resorption / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D004398:Dyneins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D051379:Mice; D008870:Microtubules / Q000378:metabolism; D008969:Molecular Sequence Data; D010010:Osteoclasts / Q000378:metabolism* / Q000473:pathology; D010012:Osteogenesis; D011485:Protein Binding; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D022142:Secretory Vesicles / Q000378:metabolism; D020692:rab3 GTP-Binding Proteins / Q000737:chemistry / Q000378:metabolism*
C533376:DYNLT1 protein, human; C533386:Dynlt1b protein, mouse; D034741:RNA, Small Interfering; D006160:Guanosine Triphosphate; C116711:Rab3d protein, mouse; D004398:Dyneins; D020692:rab3 GTP-Binding Proteins
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