Rabconnectin-3 is a functional regulator of mammalian Notch signaling.
The Notch signaling pathway is important for cell fate decisions in embryonic development and adult life. Defining the functional importance of the Notch pathway in these contexts requires the elucidation of essential signal transduction components that have not been fully characterized. Here, we show that Rabconnectin-3B is required for the Notch pathway in mammalian cells. siRNA-mediated silencing of Rabconnectin-3B in mammalian cells attenuated Notch signaling and disrupted the activation and nuclear accumulation of the Notch target Hes1. Rabconnectin-3B knockdown also disrupted V-ATPase activity in mammalian cells, consistent with previous observations in Drosophila. Pharmacological inhibition of the V-ATPase complex significantly reduced Notch signaling in mammalian cells. Finally, Rabconnectin-3B knockdown phenocopied functional disruption of Notch signaling during osteoclast differentiation. Collectively, these findings define an important role for Rabconnectin-3 and V-ATPase activity in the Notch signaling pathway in mammalian cells.
Sethi
Nilay
N
Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA.
Yan
Yan
Y
Quek
Debra
D
Schupbach
Trudi
T
Kang
Yibin
Y
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D045744:Cell Line, Tumor; D004331:Drosophila melanogaster; D055785:Gene Knockdown Techniques; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D010010:Osteoclasts / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D051880:Receptors, Notch / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D000072056:Transcription Factor HES-1; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D051792:Basic Helix-Loop-Helix Proteins; D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; D034741:RNA, Small Interfering; D051880:Receptors, Notch; D000072056:Transcription Factor HES-1; D025262:Vacuolar Proton-Translocating ATPases; C452848:DMXL2 protein, human; C489734:Hes1 protein, mouse; C078493:HES1 protein, human
EMBO Rep. 2003 Dec;4(12):1163-8
14608370
Nat Cell Biol. 2001 Apr;3(4):384-91
11283612
J Biol Chem. 2008 Mar 7;283(10):6509-18
18156632
J Biol Chem. 2002 Apr 19;277(16):13831-9
11844802
Blood. 2003 Mar 15;101(6):2227-34
12411305
Development. 2010 Jun;137(11):1825-32
20460366
Nat Genet. 2003 Mar;33(3):416-21
12590261
Curr Biol. 2004 Dec 29;14(24):2237-44
15620650
Dev Cell. 2009 Sep;17(3):387-402
19758563
Cancer Sci. 2009 Aug;100(8):1460-7
19459857
Biochem Biophys Res Commun. 2009 May 1;382(2):451-6
19289106
J Cell Biol. 2004 Jul 5;166(1):73-83
15240571
Mol Cell Biol. 2008 Oct;28(20):6402-12
18710934
Curr Opin Genet Dev. 2009 Aug;19(4):323-8
19447603
Oncogene. 2008 Sep 1;27(38):5124-31
18758481
J Cell Biol. 2009 Mar 9;184(5):621-9
19255248
Cancer Cell. 2005 Jul;8(1):13-23
16023595
Cell. 2009 Apr 17;137(2):216-33
19379690
Curr Opin Cell Biol. 2008 Apr;20(2):171-9
18342499
Science. 1999 Apr 30;284(5415):770-6
10221902
Cancer Cell. 2009 Jul 7;16(1):55-66
19573812
J Clin Invest. 1996 Feb 1;97(3):761-8
8609233
Dev Cell. 2009 May;16(5):633-47
19460341
Ann N Y Acad Sci. 2010 Mar;1192:257-68
20392245
J Cell Biol. 2008 Feb 25;180(4):755-62
18299346
Development. 2004 Mar;131(5):965-73
14973298
Cancer Cell. 2003 Jun;3(6):537-49
12842083