Download PDF
Thromb Haemost 2008; 100(05): 847-856
DOI: 10.1160/TH08-06-0351
Platelets and Blood Cells
Schattauer GmbH

Characterization of calcium- and integrin-binding protein 1 (CIB1) knockout platelets: Potential compensation by CIB family members

Jan C. DeNofrio   *
1   Curriculum in Genetics and Molecular Biology
,
Weiping Yuan   *
2   Department of Biochemistry and Biophysics
,
Brenda R. Temple
2   Department of Biochemistry and Biophysics
,
Holly R. Gentry
2   Department of Biochemistry and Biophysics
,
Leslie V. Parise
1   Curriculum in Genetics and Molecular Biology
2   Department of Biochemistry and Biophysics
3   Department of Pharmacology
4   Lineberger Comprehensive Cancer Center
5   Carolina Cardiovascular Biology Center, The University of North Carolina, Chapel Hill, North Carolina, USA
› Author AffiliationsFootnote: Research for this publication was preformed at the University of North Carolina at Chapel Hill and supported from the National Institution of Health grants 2-P01-HL45100.
Further Information

Publication History

Received 03 June 2008

Accepted after minor revision 24 August 2008

Publication Date:
22 November 2017 (online)

  PDF Download  Permissions and Reprints

Summary

Platelet aggregation requires activation of the αIIbβ3 integrin,an event regulated by the integrin cytoplasmic tails. CIB1 binds to the cytoplasmic tail of the integrin αIIb subunit. Previous overexpression and knockdown studies in murine megakaryocytes demonstrated that CIB1 inhibits integrin αIIbβ3 activation.Here we analyzed Cib1-/- mice to determine the function of CIB1 in platelets in vitro and in vivo. We found that although these mice had no overt platelet phenotype, mRNA level of CIB1 homolog CIB3 was increased in Cib1-/- megakaryocytes. In vitro binding experiments showed that recombinant CIB1, -2 and -3 bound specifically to an αIIb cytoplasmic tail peptide. Subsequent protein modeling experiments indicated that CIBs 1–3 each have a highly conserved hydrophobic binding pocket. Therefore, the potential exists for compensation for the loss of CIB1 by these CIB family members, thereby preventing pathologic thrombus formation in Cib1-/- mice.

Keywords

CIB1 - platelet - integrin αIIbβ3 activation - aggregation - knockout mouse - compensation

* Both authors contributed equally to this work.


 

  • References

  • 1 Ginsberg MH, Partridge A, Shattil SJ. Integrin regulation. Curr Opin Cell Biol 2005; 17: 509-516.
    CrossrefPubMedGoogle Scholar
  • 2 Kieffer N, Phillips DR. Platelet membrane glycoproteins: functions in cellular interactions. Annu Rev Cell Biol 1990; 06: 329-357.
    CrossrefPubMedGoogle Scholar
  • 3 Phillips DR, Charo IF, Scarborough RM. GPIIbIIIa: the responsive integrin. Cell 1991; 65: 359-362.
    CrossrefPubMedGoogle Scholar
  • 4 Andrews RK, Berndt MC. Platelet physiology and thrombosis. Thromb Res 2004; 114: 447-453.
    CrossrefPubMedGoogle Scholar
  • 5 Naik UP, Patel PM, Parise LV. Identification of a novel calcium-binding protein that interacts with the integrin alphaIIb cytoplasmic domain. J Biol Chem 1997; 272: 4651-4654.
    CrossrefPubMedGoogle Scholar
  • 6 Gentry HR, Singer AU, Betts L. et al. Structural and biochemical characterization of CIB1 delineates a new family of EF-hand-containing proteins. J Biol Chem 2005; 280: 8407-8415.
    CrossrefPubMedGoogle Scholar
  • 7 Yamniuk AP, Vogel HJ. Calcium- and magnesium-dependent interactions between calcium- and integrin binding protein and the integrin alphaIIb cytoplasmic domain. Protein Sci 2005; 14: 1429-1437.
    PubMedGoogle Scholar
  • 8 Wu X, Lieber MR. Interaction between DNA-dependent protein kinase and a novel protein, KIP. Mutat Res 1997; 385: 13-20.
    CrossrefPubMedGoogle Scholar
  • 9 Kauselmann G, Weiler M, Wulff P. et al. The pololike protein kinases Fnk and Snk associate with a Ca(2+)- and integrin-binding protein and are regulated dynamically with synaptic plasticity. EMBO J 1999; 18: 5528-5539.
    CrossrefPubMedGoogle Scholar
  • 10 Leisner TM, Liu M, Jaffer ZM. et al. Essential role of CIB1 in regulating PAK1 activation and cell migration. J Cell Biol 2005; 170: 465-476.
    CrossrefPubMedGoogle Scholar
  • 11 Stabler SM, Ostrowski LL, Janicki SM. et al. A myristoylated calcium-binding protein that preferentially interacts with the Alzheimer’s disease presenilin 2 protein. J Cell Biol 1999; 145: 1277-1292.
    CrossrefPubMedGoogle Scholar
  • 12 Naik MU, Naik UP. Calcium-and integrin-binding protein regulates focal adhesion kinase activity during platelet spreading on immobilized fibrinogen. Blood 2003; 102: 3629-3636.
    CrossrefPubMedGoogle Scholar
  • 13 White C, Yang J, Monteiro MJ. et al. CIB1, a ubiquitously expressed Ca2+-binding protein ligand of the InsP3 receptor Ca2+ release channel. J Biol Chem 2006; 281: 20825-20833.
    CrossrefPubMedGoogle Scholar
  • 14 Zayed MA, Yuan W, Leisner TM. et al. Calcium and integrin-binding protein 1 regulates endothelial cells and ischemia-induced pathological and adaptive angiogenesis. Circ Res 2007; 101: 1185-1193.
    CrossrefPubMedGoogle Scholar
  • 15 Yuan W, Leisner TM, McFadden AW. et al. CIB1 is essential for mouse spermatogenesis. Mol Cell Biol 2006; 26: 8507-8514.
    CrossrefPubMedGoogle Scholar
  • 16 Yuan W, Leisner TM, McFadden AW. et al. CIB1 is an endogenous inhibitor of agonist-induced integrin alphaIIbbeta3 activation. J Cell Biol 2006; 172: 169-175.
    CrossrefPubMedGoogle Scholar
  • 17 Naik UP, Naik MU. Association of CIB with GPIIb/ IIIa during outside-in signaling is required for platelet spreading on fibrinogen. Blood 2003; 102: 1355-1362.
    CrossrefPubMedGoogle Scholar
  • 18 Shiraga M, Ritchie A, Aidoudi S. et al. Primary megakaryocytes reveal a role for transcription factor NF-E2 in integrin alpha IIb beta 3 signaling. J Cell Biol 1999; 147: 1419-1430.
    CrossrefPubMedGoogle Scholar
  • 19 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25: 402-408.
    CrossrefPubMedGoogle Scholar
  • 20 Schefe JH, Lehmann KE, Buschmann IR. et al. Quantitative real-time RT-PCR data analysis: current concepts and the novel “gene expression’s CT difference” formula. J Mol Med 2006; 84: 901-910.
    CrossrefPubMedGoogle Scholar
  • 21 Bairoch A, Apweiler R, Wu CH. et al. The Universal Protein Resource (UniProt). Nucleic Acids Res 2005; 33 Database issue D154-D159.
    PubMedGoogle Scholar
  • 22 Altschul SF, Lipman DJ. Protein database searches for multiple alignments. Proc Natl Acad Sci USA 1990; 87: 5509-5513.
    CrossrefPubMedGoogle Scholar
  • 23 Chenna R, Sugawara H, Koike T. et al. Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res 2003; 31: 3497-3500.
    CrossrefPubMedGoogle Scholar
  • 24 Subramaniam M, Frenette PS, Saffaripour S. et al. Defects in hemostasis in P-selectin-deficient mice. Blood 1996; 87: 1238-1242.
    PubMedGoogle Scholar
  • 25 Denis CV, Wagner DD. Platelet adhesion receptors and their ligands in mouse models of thrombosis. Arterioscler Thromb Vasc Biol 2007; 27: 728-739.
    CrossrefPubMedGoogle Scholar
  • 26 Furie B, Furie BC. Thrombus formation in vivo. J Clin Invest 2005; 115: 3355-3362.
    CrossrefPubMedGoogle Scholar
  • 27 Kurz KD, Main BW, Sandusky GE. Rat model of arterial thrombosis induced by ferric chloride. Thromb Res 1990; 60: 269-280.
    CrossrefPubMedGoogle Scholar
  • 28 Tsuboi S. Calcium integrin-binding protein activates platelet integrin alphaIIbbeta 3. J Biol Chem 2002; 277: 1919-1923.
    CrossrefPubMedGoogle Scholar
  • 29 Mayhew MW, Webb DJ, Kovalenko M. et al. Identification of protein networks associated with the PAK1-betaPIX-GIT1-paxillin signaling complex by mass spectrometry. J Proteome Res 2006; 05: 2417-2423.
    CrossrefPubMedGoogle Scholar
  • 30 Barry WT, Boudignon-Proudhon C, Shock DD. et al. Molecular basis of CIB binding to the integrin alpha IIb cytoplasmic domain. J Biol Chem 2002; 277: 28877-28883.
    CrossrefPubMedGoogle Scholar
  • 31 Shock DD, Naik UP, Brittain JE. et al. Calcium-dependent properties of CIB binding to the integrin alphaIIb cytoplasmic domain and translocation to the platelet cytoskeleton. Biochem J 1999; 342: 729-735.
    CrossrefPubMedGoogle Scholar
  • 32 Yamniuk AP, Ishida H, Vogel HJ. The interaction between calcium- and integrin-binding protein 1 and the alphaIIb integrin cytoplasmic domain involves a novel C-terminal displacement mechanism. J Biol Chem 2006; 281: 26455-26464.
    CrossrefPubMedGoogle Scholar
  • 33 Han J, Lim CJ, Watanabe N. et al. Reconstructing and deconstructing agonist-induced activation of integrin alphaIIbbeta3. Curr Biol 2006; 16: 1796-1806.
    CrossrefPubMedGoogle Scholar
  • 34 Petrich BG, Marchese P, Ruggeri ZM. et al. Talin is required for integrin-mediated platelet function in hemostasis and thrombosis. J Exp Med 2007; 204: 3103-3111.
    CrossrefPubMedGoogle Scholar
  • 35 Petrich BG, Fogelstrand P, Partridge AW. et al. The antithrombotic potential of selective blockade of talindependent integrin alpha IIb beta 3 (platelet GPIIbIIIa) activation. J Clin Invest 2007; 117: 2250-2259.
    CrossrefPubMedGoogle Scholar
  • 36 Leisner TM, Yuan W, Denofrio JC. et al. Tickling the tails: cytoplasmic domain proteins that regulate integrin alphaIIbbeta3 activation. Curr Opin Hematol 2007; 14: 255-261.
    CrossrefPubMedGoogle Scholar
  • 37 Larkin D, Murphy D, Reilly DF. et al. ICln, a novel integrin alphaIIbbeta3-associated protein, functionally regulates platelet activation. J Biol Chem 2004; 279: 27286-27293.
    CrossrefPubMedGoogle Scholar
  • 38 Leung-Hagesteijn CY, Milankov K, Michalak M. et al. Cell attachment to extracellular matrix substrates is inhibited upon downregulation of expression of calreticulin, an intracellular integrin alpha-subunit-binding protein. J Cell Sci 1994; 107: 589-600.
    PubMedGoogle Scholar
  • 39 Liu QY, Corjay M, Feuerstein GZ. et al. Identification and characterization of triosephosphate isomerase that specifically interacts with the integrin alphaIIb cytoplasmic domain. Biochem Pharmacol 2006; 72: 551-557.
    CrossrefPubMedGoogle Scholar
  • 40 Reilly D, Larkin D, Devocelle M. et al. Calreticulin-independent regulation of the platelet integrin alphaIIbbeta3 by the KVGFFKR alphaIIb-cytoplasmic motif. Platelets 2004; 15: 43-54.
    CrossrefPubMedGoogle Scholar
  • 41 Vijayan KV, Liu Y, Li TT. et al. Protein phosphatase 1 associates with the integrin alphaIIb subunit and regulates signaling. J Biol Chem 2004; 279: 33039-33042.
    CrossrefPubMedGoogle Scholar
  • 42 Liu J, DeNofrio J, Yuan W. et al. Genetic Manipulation of Megakaryocytes to Study Platelet Function. In: Gerald PS. Current Topics in Developmental Biology. Academic Press; 2007: 311-335.
    Google Scholar
  • 43 Vallar L, Melchior C, Plancon S. et al. Divalent cations differentially regulate integrin alphaIIb cytoplasmic tail binding to beta3 and to calcium- and integrin-binding protein. J Biol Chem 1999; 274: 17257-17266.
    CrossrefPubMedGoogle Scholar
  • 44 Pearce AC, Senis YA, Billadeau DD. et al. Vav1 and vav3 have critical but redundant roles in mediating platelet activation by collagen. J Biol Chem 2004; 279: 53955-53962.
    CrossrefPubMedGoogle Scholar
  • 45 Schraw TD, Crawford GL, Ren Q. et al. Platelets from Munc18c heterozygous mice exhibit normal stimulus-induced release. Thromb Haemost 2004; 92: 829-837.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 46 Pearce AC, McCarty OJ, Calaminus SD. et al. Vav family proteins are required for optimal regulation of PLCgamma2 by integrin alphaIIbbeta3. Biochem J 2007; 401: 753-761.
    CrossrefPubMedGoogle Scholar
  • 47 Pearce AC, Wilde JI, Doody GM. et al. Vav1, but not Vav2, contributes to platelet aggregation by CRP and thrombin, but neither is required for regulation of phospholipase C. Blood 2002; 100: 3561-3569.
    CrossrefPubMedGoogle Scholar