C-terminal region of Mad2 plays an important role during mitotic spindle checkpoint in fission yeast Schizosaccharomyces pombe.
The mitotic arrest deficiency 2 (Mad2) protein is an essential component of the spindle assembly checkpoint that interacts with Cdc20/Slp1 and inhibit its ability to activate anaphase promoting complex/cyclosome (APC/C). In bladder cancer cell line the C-terminal residue of the mad2 gene has been found to be deleted. In this study we tried to understand the role of the C-terminal region of mad2 on the spindle checkpoint function. To envisage the role of C-terminal region of Mad2, we truncated 25 residues of Mad2 C-terminal region in fission yeast S.pombe and characterized its effect on spindle assembly checkpoint function. The cells containing C-terminal truncation of Mad2 exhibit sensitivity towards microtubule destabilizing agent suggesting perturbation of spindle assembly checkpoint. Further, the C-terminal truncation of Mad2 exhibit reduced viability in the nda3-KM311 mutant background at non-permissive temperature. Truncation in mad2 gene also affects its foci forming ability at unattached kinetochore suggesting that the mad2-∆CT mutant is unable to maintain spindle checkpoint activation. However, in response to the defective microtubule, only brief delay of mitotic progression was observed in Mad2 C-terminal truncation mutant. In addition we have shown that the deletion of two β strands of Mad2 protein abolishes its ability to interact with APC activator protein Slp1/Cdc20. We purpose that the truncation of two β strands (β7 and β8) of Mad2 destabilize the safety belt and affect the Cdc20-Mad2 interaction leading to defects in the spindle checkpoint activation.
Molecular and Structural Biology Division, Central Drug Research Institute (CSIR), Sector 10, Jankipuram Extension, Sitapur Road, Lucknow, 226031, India.
Gaurav Kumar
GK
Singh
Molecular and Structural Biology Division, Central Drug Research Institute (CSIR), Sector 10, Jankipuram Extension, Sitapur Road, Lucknow, 226031, India.
Sharanbasappa Shrimant
SS
Karade
Molecular and Structural Biology Division, Central Drug Research Institute (CSIR), Sector 10, Jankipuram Extension, Sitapur Road, Lucknow, 226031, India.
Rajeev
R
Ranjan
Molecular and Structural Biology Division, Central Drug Research Institute (CSIR), Sector 10, Jankipuram Extension, Sitapur Road, Lucknow, 226031, India.
Nafees
N
Ahamad
Molecular and Structural Biology Division, Central Drug Research Institute (CSIR), Sector 10, Jankipuram Extension, Sitapur Road, Lucknow, 226031, India. shakil_ahmed@cdri.res.in.
Shakil
S
Ahmed
D064199:Cdc20 Proteins / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D059566:M Phase Cell Cycle Checkpoints; D064246:Mad2 Proteins / Q000737:chemistry; D008938:Mitosis; D008958:Models, Molecular; D017433:Protein Structure, Secondary; D012568:Schizosaccharomyces / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000072020:Synthetic Lethal Mutations
D064199:Cdc20 Proteins; C577498:Cdc20 protein, S pombe; D018797:Cell Cycle Proteins; C577697:MAD2L1 protein, human; D064246:Mad2 Proteins; D029702:Schizosaccharomyces pombe Proteins
Mad2; Nda3; Slp1/Cdc20; Spindle assembly checkpoint
EMBO J. 2002 May 15;21(10):2496-506
12006501
Cancer Cell. 2007 Jan;11(1):9-23
17189715
PLoS One. 2014 Feb 21;9(2):e89587
24586893
Nat Rev Mol Cell Biol. 2007 May;8(5):379-93
17426725
Int J Cancer. 2001 Jul 20;95(4):223-7
11400114
Cell Cycle. 2013 Mar 15;12 (6):961-71
23442800
Yeast. 1998 Jul;14 (10 ):943-51
9717240
Biochim Biophys Acta. 2016 Feb;1863(2):189-97
26581324
Cell. 1991 Aug 9;66(3):519-31
1651172
Mol Cell. 2007 Jul 6;27(1):3-16
17612486
Curr Biol. 2015 Oct 19;25(20):R1002-18
26485365
J Cell Sci. 2012 Oct 15;125(Pt 20):4720-7
22825872
Nat Struct Biol. 2000 Mar;7(3):224-9
10700282
Cell. 1991 Aug 9;66(3):507-17
1651171
PLoS Biol. 2008 Mar 4;6(3):e50
18318601
Cell. 1984 Dec;39(2 Pt 1):349-58
6094012
Nucleic Acids Res. 2014 Jul;42(Web Server issue):W320-4
24753421
Curr Protoc Bioinformatics. 2006 Oct;Chapter 5:Unit-5.6
18428767
Nat Struct Mol Biol. 2004 Apr;11(4):338-45
15024386
Curr Biol. 2005 Feb 8;15(3):214-25
15694304
Oncogene. 2008 Jan 10;27(3):347-57
17621272
Science. 1996 Oct 11;274(5285):246-8
8824189
Curr Opin Genet Dev. 1999 Feb;9(1):69-75
10072359
Nature. 2012 Mar 21;484(7393):208-13
22437499
Nat Rev Mol Cell Biol. 2006 Sep;7(9):644-56
16896351
J Cell Biol. 1996 Jan;132(1-2):111-23
8567717
Methods Enzymol. 1991;194:795-823
2005825
Cell. 2007 Nov 16;131(4):730-43
18022367
Mol Cell. 2008 Nov 21;32(4):576-83
19026787
Cell. 2003 Feb 21;112(4):407-21
12600307
Genet Res (Camb). 2011 Aug;93(4):275-83
21767457
Curr Biol. 2012 Feb 7;22(3):180-90
22209528
Mol Cell. 2002 Jan;9(1):59-71
11804586