Targeted Mutagenesis of the Angiogenic Protein CCN1 (CYR61)
Open accessAbstract
The matricellular protein CCN1 (CYR61) regulates multiple cellular processes and plays essential roles in embryonic vascular development. A ligand of several integrin receptors, CCN1 acts through integrin alpha6beta1 and heparan sulfate proteoglycans (HSPGs) to promote specific functions in fibroblasts, smooth muscle cells, and endothelial cells. We have previously identified a novel alpha6beta1 binding site, T1, in domain III of CCN1. Here we uncover two novel 16-residue sequences, H1 and H2, in domain IV that can support alpha6beta1- and HSPGs-dependent cell adhesion, suggesting that these sequences contain closely juxtaposed or overlapping sites for interaction with alpha6beta1 and HSPGs. Furthermore, fibroblast adhesion to the H1 and H2 peptides is sufficient to induce prolonged MAPK activation, whereas adhesion to T1 induces transient MAPK activation. To dissect the roles of these sites in CCN1 function, we have created mutants disrupted in T1, H1, and H2 or in all three sites in the context of full-length CCN1. We show that the T1 and H1/H2 sites are functionally non-equivalent, and disruption of these sites differentially affected cell adhesion, migration, mitogen-activated protein kinase activation, and regulation of gene expression. Disruption of all three sites completely abolished alpha6beta1-HSPG-mediated cellular activities. All mutants disrupting T1, H1, and H2 fully retain alphavbeta3-mediated pro-angiogenic activities, indicating that these mutants are biologically active and are defective only in alpha6beta1-HSPG-mediated functions. Together, these findings identify and dissect the differential roles of the three sites (T1, H1, H2) required for alpha6beta1-HSPG-dependent CCN1 activities and provide a strategy to investigate these alpha6beta1-HSPG-specific activities in vivo.
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CCN1 lacks the canonical RGD sequence that binds many integrins; instead, specific non-canonical binding sites for several integrins have been identified.
“CCN1 does not contain the canonical RGD sequence that can bind a number of integrins [54]. Instead, specific non-canonical binding sites of CCN1 for several integrins have been identified (Fig. 1) [63–66].”
From CCN1/CYR61: the very model of a modern matricellular protein (2011) · cites this paper as [66]
Distinct integrin-binding sites of CCN1 can function independently: disrupting α6β1-binding sites abolishes α6β1-dependent activities without affecting αvβ3-mediated functions, and disrupting the αvβ3-binding site abolishes αvβ3- but not α6β1-mediated activities.
“CCN1 mutants that disrupt its α6β1-binding sites abolish α6β1-dependent activities without affecting αvβ3-mediated functions [66]. Likewise, disruption of the CCN1 αvβ3-binding site abolishes αvβ3-but not α6β1-mediated activities [65].”
From CCN1/CYR61: the very model of a modern matricellular protein (2011) · cites this paper as [66]
A short CCN1 peptide containing the α6β1-HSPG binding sites is sufficient to support cell adhesion and recapitulate sustained ERK activation by immobilized CCN1.
“A short CCN1 peptide containing the α6β1–HSPG binding sites is sufficient to support cell adhesion and recapitulate the sustained activation of extracellular signal-regulated kinase (ERK) by immobilized CCN1 [66].”
From CCN1/CYR61: the very model of a modern matricellular protein (2011) · cites this paper as [66]
Cites
Marshall, C. J. (1995) · Cell · doi:10.1016/0092-8674(95)90401-8 · Cited by 4,557 · see citing works
The PSIPRED protein structure prediction server
McGuffin, L. J., Bryson, K., & Jones, D. T. (2000) · Bioinformatics · doi:10.1093/bioinformatics/16.4.404 · Cited by 3,841 · see citing works
Matricellular proteins: extracellular modulators of cell function
BORNSTEIN, P. K., & Sage, E. H. (2002) · Current Opinion in Cell Biology · doi:10.1016/s0955-0674(02)00361-7 · Cited by 966 · see citing works
A 48-well micro chemotaxis assembly for rapid and accurate measurement of leukocyte migration
Falk, W., Goodwin, R. H., & Leonard, E. J. (1980) · Journal of Immunological Methods · doi:10.1016/0022-1759(80)90211-2 · Cited by 942 · see citing works
Molecular interpretation of ERK signal duration by immediate early gene products
Murphy, L. O., Smith, S. W., Chen, R., Fingar, D. C., & Blenis, J. (2002) · Nature Cell Biology · doi:10.1038/ncb822 · Cited by 928 · see citing works
Sensing the environment: a historical perspective on integrin signal transduction
Miranti, C. K., & Brugge, J. S. (2002) · Nature Cell Biology · doi:10.1038/ncb0402-e83 · Cited by 851 · see citing works
Isberg, R. R., & Leong, J. M. (1990) · Cell · doi:10.1016/0092-8674(90)90099-z · Cited by 798 · see citing works
Ivković, S., Yoon, B. S., Popoff, S. N., Safadi, F. F., Libuda, D. E., Stephenson, R. C., Daluiski, A., & Lyons, K. M. (2003) · Development · doi:10.1242/dev.00505 · Cited by 705 · see citing works
Diversity of function is inherent in matricellular proteins: an appraisal of thrombospondin 1.
Bornstein, P. (1995) · The Journal of cell biology · doi:10.1083/jcb.130.3.503 · Cited by 666 · see citing works
The CCN Family of Angiogenic Regulators: The Integrin Connection
Lau, L. F., & Lam, S. C.-T. (1999) · Experimental Cell Research · doi:10.1006/excr.1999.4456 · Cited by 645 · see citing works
Self-citationPeer Bork, B. (1993) · FEBS Letters · doi:10.1016/0014-5793(93)80155-n · Cited by 623 · see citing works
The Connective Tissue Growth Factor/Cysteine- Rich 61/Nephroblastoma Overexpressed (CCN) Family
Brigstock, D. R. (1999) · Endocrine Reviews · doi:10.1210/edrv.20.2.0360 · Cited by 588 · see citing works
A 48-well micro chemotaxis assembly for rapid and accurate measurement of leukocyte migration
Falk, W., Goodwin, R. H., & Leonard, E. J. (1980) · Journal of Immunological Methods · doi:10.1016/s0022-1759(80)80014-7 · Cited by 585 · see citing works
Babic, A. M., Kireeva, M. L., Kolesnikova, T. V., & Lau, L. F. (1998) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.95.11.6355 · Cited by 495 · see citing works
Self-citationMouse egg integrin α6β1functions as a sperm receptor
Almeida, E. A. C., Huovila, A.-P. J., Sutherland, A. E., Stephens, L., Calarco, P. G., Shaw§, L. M., Mercurio, A. M., Sonnenberg, A., Primakoff, P., Myles, D. G., et al. (1995) · Cell · doi:10.1016/s0092-8674(05)80014-5 · Cited by 470 · see citing works
CYR61 (CCN1) Is Essential for Placental Development and Vascular Integrity
Mo, F.-E., Muntean, A. G., Chen, C.-C., Stolz, D. B., Watkins, S. C., & Lau, L. F. (2002) · Molecular and Cellular Biology · doi:10.1128/mcb.22.24.8709-8720.2002 · Cited by 416 · see citing works
Self-citationNOV (nephroblastoma overexpressed) and the CCN family of genes: structural and functional issues
Perbal, B. (2001) · Molecular Pathology · doi:10.1136/mp.54.2.57 · Cited by 345 · see citing works
A small-molecule inhibitor of skeletal muscle myosin II
Cheung, A., Dantzig, J. A., Hollingworth, S., Baylor, S. M., Goldman, Y. E., Mitchison, T. J., & Straight, A. F. (2001) · Nature Cell Biology · doi:10.1038/ncb734 · Cited by 342 · see citing works
Kireeva, M. L., Mo, F.-E., Yang, G. P., & Lau, L. F. (1996) · Molecular and Cellular Biology · doi:10.1128/mcb.16.4.1326 · Cited by 341 · see citing works
Self-citationExpression of cyr61, a growth factor-inducible immediate-early gene.
O'Brien, T. P., Yang, G. P., Sanders, L., & Lau, L. F. (1990) · Molecular and Cellular Biology · doi:10.1128/mcb.10.7.3569 · Cited by 310 · see citing works
Self-citationChen, C.-C., Chen, N., & Lau, L. F. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m008087200 · Cited by 305 · see citing works
Self-citationThe Angiogenic Factor Cyr61 Activates a Genetic Program for Wound Healing in Human Skin Fibroblasts
Chen, C.-C., Mo, F.-E., & Lau, L. F. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m107666200 · Cited by 300 · see citing works
Self-citationThrombospondins 1 and 2 function as inhibitors of angiogenesis
Armstrong, L. C., & BORNSTEIN, P. K. (2003) · Matrix Biology · doi:10.1016/s0945-053x(03)00005-2 · Cited by 287 · see citing works
Tam, E. M., Morrison, C., Wu, Y., Stack, M. S., & Overall, C. M. (2004) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0305862101 · Cited by 277 · see citing works
Mutations in the CCN gene family member WISP3 cause progressive pseudorheumatoid dysplasia
Hurvitz, J., Suwairi, W. M., Hul, W. V., El‐Shanti, H., Superti‐Furga, A., Roudier, J., Holderbaum, D. F., Pauli, R. M., Herd, J. K., Hul, E. V., et al. (1999) · Nature Genetics · doi:10.1038/12699 · Cited by 271 · see citing works
Crystal structure of the TSP-1 type 1 repeats
Tan, K., Duquette, M. A., Liu, J., Dong, Y., Zhang, R., Joachimiak, A., Lawler, J., & Wang, J. (2002) · The Journal of Cell Biology · doi:10.1083/jcb.200206062 · Cited by 258 · see citing works
Leu, S.-J., Lam, S. C.-T., & Lau, L. F. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m209288200 · Cited by 247 · see citing works
Self-citationSchober, J. M., Chen, N., Grzeszkiewicz, T. M., Jovanovic, I., Emeson, E. E., Ugarova, T. P., Ye, R. D., Lau, L. F., & Lam, S. C.-T. (2002) · Blood · doi:10.1182/blood.v99.12.4457 · Cited by 242 · see citing works
Self-citationTwo RGD-independent αvβ3 Integrin Binding Sites on Tumstatin Regulate Distinct Anti-tumor Properties
Maeshima, Y., Colorado, P. C., & Kalluri, R. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.c000186200 · Cited by 229 · see citing works
The α6β1 (VLA-6) and α6β4 protein complexes: tissue distribution and biochemical properties
Sonnenberg, A., Linders, C. J. T., Daams, J. H., et al. (1990) · Journal of Cell Science · doi:10.1242/jcs.96.2.207 · Cited by 217 · see citing works
Jedsadayanmata, A., Chen, C.-C., Kireeva, M. L., Lau, L. F., & Lam, S. C.-T. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.34.24321 · Cited by 204 · see citing works
Self-citationKireeva, M. L., Lam, S. C.-T., & Lau, L. F. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.5.3090 · Cited by 203 · see citing works
Self-citationIntegrin-specific signaling pathways controlling focal adhesion formation and cell migration
Mostafavi‐Pour, Z., Askari, J. A., Parkinson, S. J., et al. (2003) · The Journal of Cell Biology · doi:10.1083/jcb.200210176 · Cited by 203 · see citing works
Sakamoto, K., Yamaguchi, S., Ando, R., Miyawaki, A., Kabasawa, Y., Takagi, M., Li, C. L., Perbal, B., & Katsube, K. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m203727200 · Cited by 202 · see citing works
Chen, N., Chen, C.-C., & Lau, L. F. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m003040200 · Cited by 195 · see citing works
Self-citationYang, G. P., & Lau, L. F. (1991) · PubMed · Cited by 187 · see citing works
Self-citationGrzeszkiewicz, T. M., Kirschling, D. J., Chen, N., & Lau, L. F. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m100978200 · Cited by 162 · see citing works
Self-citationCCN3 (NOV) Interacts with Connexin43 in C6 Glioma Cells
Fu, C. T., Bechberger, J. F., Ozog, M. A., Perbal, B., & Naus, C. C. G. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m403952200 · Cited by 155 · see citing works
Wong, M., Kireeva, M. L., Kolesnikova, T. V., & Lau, L. F. (1997) · Developmental Biology · doi:10.1006/dbio.1997.8766 · Cited by 155 · see citing works
Self-citationPerbal, B., Martinerie, C., Sainson, R. C. A., Werner, M., He, B., & Roizman, B. (1999) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.96.3.869 · Cited by 146 · see citing works
Predicting novel protein folds by using FRAGFOLD
Jones, D. T. (2001) · Proteins Structure Function and Bioinformatics · doi:10.1002/prot.1171 · Cited by 141 · see citing works
O'Brien, T. P., & Lau, L. F. (1992) · PubMed · Cited by 140 · see citing works
Self-citationChen, N., Leu, S.-J., Todorović, V., Lam, S. C.-T., & Lau, L. F. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m406813200 · Cited by 132 · see citing works
Self-citationIdentification of a novel integrin α6β1 binding site in the angiogenic Inducer CCN1 (CYR61)
Leu, S.-J., Liu, Y., Chen, N., Chen, C.-C., Lam, S. C.-T., & Lau, L. F. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m305862200 · Cited by 131 · see citing works
Self-citationRecombinant Laminin-8 (α4β1γ1)
Kortesmaa, J., Yurchenco, P. D., & Tryggvason, K. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.275.20.14853 · Cited by 127 · see citing works
The angiogenic factor CYR61 in breast cancer: molecular pathology and therapeutic perspectives.
Men√©ndez, J. A., Mehmi, I., Griggs, D. W., & Lupu, R. (2003) · Endocrine Related Cancer · doi:10.1677/erc.0.0100141 · Cited by 120 · see citing works
Hilfiker, A., Hilfiker-Kleiner, D., Fuchs, M., Kaminski, K., Lichtenberg, A., Rothkötter, H.-J., Schieffer, B., & Drexler, H. (2002) · Circulation · doi:10.1161/01.cir.0000021426.87274.62 · Cited by 114 · see citing works
Recognition of the N-terminal Modules of Thrombospondin-1 and Thrombospondin-2 by α6β1 Integrin
Calzada, M. J., Sipes, J. M., Krutzsch, H. C., et al. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m302014200 · Cited by 103 · see citing works
Ball, D. K., Surveyor, G. A., Diehl, J. R., Steffen, C. L., Uzumcu, M., Mirando, M. A., & Brigstock, D. R. (1998) · Biology of Reproduction · doi:10.1095/biolreprod59.4.828 · Cited by 96 · see citing works
Schober, J. M., Lau, L. F., Ugarova, T. P., & Lam, S. C.-T. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m301534200 · Cited by 76 · see citing works
Self-citationLatinkic, B. V. (2001) · Endocrinology · doi:10.1210/en.142.6.2549 · Cited by 61 · see citing works
Stimulation of Angiogenesis by Cyr61 Gene: A New Therapeutic Candidate
Fataccioli, V., Abergel, V., Wingertsmann, L., Neuville, P., Spitz, E., Adnot, S., Calenda, V., & Teiger, E. (2002) · Human Gene Therapy · doi:10.1089/10430340260185094 · Cited by 58 · see citing works
Acuña-Cueva, E. R., Hueso‐Ureña, F., Jiménez‐Pulido, S. B., & Moreno‐Carretero, M. N. (2000) · Journal of Molecular Modeling · doi:10.1007/s0089400060433 · Cited by 27 · see citing works
Wu, K., Yee, A., Zhu, N. L., Gordon, E. M., & Hall, F. L. (2000) · International Journal of Molecular Medicine · doi:10.3892/ijmm.6.4.433 · Cited by 24 · see citing works
Grzeszkiewicz, T. M. (2002) · Endocrinology · doi:10.1210/en.143.4.1441 · Cited by 23 · see citing works
Similar early gene responses to ligand-activated EGFR and neu tyrosine kinases in NIH3T3 cells.
KOSKINEN, P. K., Lehväslaiho, H., Macdonald-Bravo, H., et al. (1990) · PubMed · Cited by 15 · see citing works
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Chen, C.-C., & Lau, L. F. (2009) · The International Journal of Biochemistry & Cell Biology · doi:10.1016/j.biocel.2008.07.025 · Cited by 549 · see citing works
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Lau, L. F. (2011) · Cellular and Molecular Life Sciences · doi:10.1007/s00018-011-0778-3 · Cited by 338 · see citing works
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Jun, J.-I., Kim, K.-H., & Lau, L. F. (2015) · Nature Communications · doi:10.1038/ncomms8386 · Cited by 173 · see citing works
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Chen, C., Young, J. L., Monzon, R. I., Chen, N., Todorović, V., & Lau, L. F. (2007) · The EMBO Journal · doi:10.1038/sj.emboj.7601596 · Cited by 143 · see citing works
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Todorovicç, V., Chen, C.-C., Hay, N., & Lau, L. F. (2005) · The Journal of Cell Biology · doi:10.1083/jcb.200504015 · Cited by 132 · see citing works
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Mo, F.-E., & Lau, L. F. (2006) · Circulation Research · doi:10.1161/01.res.0000248426.35019.89 · Cited by 112 · see citing works
Self-citationBorkham-Kamphorst, E., Schaffrath, C., Van de Leur, E., Haas, U., Tihaa, L., Meurer, S. K., Nevzorova, Y. A., Liedtke, C., & Weiskirchen, R. (2014) · Biochimica et Biophysica Acta (BBA) - Molecular Cell Research · doi:10.1016/j.bbamcr.2014.01.023 · Cited by 105 · see citing works
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Vellon, L., Menendez, J. A., & Lupu, R. (2005) · Oncogene · doi:10.1038/sj.onc.1208452 · Cited by 98 · see citing works
Juric, V., Chen, C.-C., & Lau, L. F. (2009) · Molecular and Cellular Biology · doi:10.1128/mcb.00064-09 · Cited by 70 · see citing works
Self-citationFranzen, C. A., Chen, C.-C., Todorović, V., Juric, V., Monzon, R. I., & Lau, L. F. (2009) · Molecular Cancer Research · doi:10.1158/1541-7786.mcr-09-0017 · Cited by 66 · see citing works
Self-citationWolf, N., Yang, W., Dunk, C. E., Gashaw, I., Lye, S. J., Ring, T., Schmidt, M., Winterhager, E., & Gellhaus, A. (2010) · Endocrinology · doi:10.1210/en.2009-1195 · Cited by 52 · see citing works
Chen, C., & Lau, L. F. (2010) · Journal of Cell Communication and Signaling · doi:10.1007/s12079-009-0080-4 · Cited by 47 · see citing works
Self-citationKassis, J. N., Virador, V. M., Guancial, E. A., Kimm, D., Ho, A. S., Mishra, M., Chuang, E. Y., Cook, J., Gius, D., & Kohn, E. C. (2009) · The Journal of Pathology · doi:10.1002/path.2557 · Cited by 34 · see citing works
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Chen, C.-C., Juric, V., & Lau, L. F. (2011) · Advances in Experimental Medicine and Biology · doi:10.1007/978-1-4419-6612-4_63 · Cited by 8 · see citing works
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- Authors
- Leu, S.-J., Chen, N., Chen, C.-C., Todorović, V., Bai, T., Juric, V., Liu, Y., Yan, G., Lam, S. C.-T., & Lau, L. F.
- Affiliations
- University of Illinois Chicago (US)
- Corresponding author
- Lau, Lester F.
- ORCID
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2004-10
- Volume
- 279
- Issue
- 42
- Pages
- 44177-44187
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2160948226
- Cited by
- 72
- References
- 56
- Citations by year
- 2026: 1 · 2025: 5 · 2024: 1 · 2023: 1 · 2022: 3 · 2021: 3 · 2020: 2 · 2019: 2 · 2018: 2 · 2017: 3 · 2016: 2 · 2015: 5 · 2014: 4 · 2013: 4 · 2012: 4
- FWCI
- 1.30
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01ca080080-05)
- National Heart, Lung, and Blood Institute (HL41793)
- National Cancer Institute (CA-80080, R01CA046565, CA-46565)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Connective Tissue Growth Factor Research
- Health Sciences › Medicine › Rheumatology › Bone and Dental Protein Studies
- Health Sciences › Medicine › Immunology and Allergy › Cell Adhesion Molecules Research
- Keywords
- CYR61, matricellular proteins, alpha6beta1 integrin, heparan sulfate proteoglycans, cell adhesion, MAPK activation, angiogenesis, integrin αvβ3, integrin binding site, fibroblast migration, gene expression regulation, targeted mutagenesis
- MeSH terms
- Amino Acid Sequence, Animals, Cell Adhesion, Enzyme-Linked Immunosorbent Assay, Fibroblasts, Humans, Molecular Sequence Data, Muscle, Smooth, Vascular, Peptide Fragments, Recombinant Fusion Proteins, Mutagenesis, Site-Directed, Immediate-Early Proteins, DNA Primers, Integrin alpha6beta1, Mice, Cysteine-Rich Protein 61
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