Functional Characterization of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) N- and C-Terminal Domains during Xenopus laevis Development

Extracellular matrix (ECM) remodeling is essential for facilitating developmental processes. ECM remodeling, accomplished by matrix metalloproteinases (MMPs), is regulated by endogenous tissue inhibitors of metalloproteinases (TIMPs). While the TIMP N-terminal domain is involved in inhibition of MMP...

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Main Authors: M. A. Nieuwesteeg, J. A. Willson, M. Cepeda, M. A. Fox, S. Damjanovski
Format: Article
Language:English
Published: Wiley 2014-01-01
Series:The Scientific World Journal
Online Access:http://dx.doi.org/10.1155/2014/467907
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author M. A. Nieuwesteeg
J. A. Willson
M. Cepeda
M. A. Fox
S. Damjanovski
author_facet M. A. Nieuwesteeg
J. A. Willson
M. Cepeda
M. A. Fox
S. Damjanovski
author_sort M. A. Nieuwesteeg
collection DOAJ
description Extracellular matrix (ECM) remodeling is essential for facilitating developmental processes. ECM remodeling, accomplished by matrix metalloproteinases (MMPs), is regulated by endogenous tissue inhibitors of metalloproteinases (TIMPs). While the TIMP N-terminal domain is involved in inhibition of MMP activity, the C-terminal domain exhibits cell-signaling activity, which is TIMP and cell type dependent. We have previously examined the distinct roles of the Xenopus laevis TIMP-2 and -3 C-terminal domains during development and here examined the unique roles of TIMP-1 N- and C-terminal domains in early X. laevis embryos. mRNA microinjection was used to overexpress full-length TIMP-1 or its individual N- or C-terminal domains in embryos. Full-length and C-terminal TIMP-1 resulted in increased lethality compared to N-terminal TIMP-1. Overexpression of C-terminal TIMP-1 resulted in significant decreases in mRNA levels of proteolytic genes including TIMP-2, RECK, MMP-2, and MMP-9, corresponding to decreases in MMP-2 and -9 protein levels, as well as decreased MMP-2 and MMP-9 activities. These trends were not observed with the N-terminus. Our research suggests that the individual domains of TIMP-1 are capable of playing distinct roles in regulating the ECM proteolytic network during development and that the unique functions of these domains are moderated in the endogenous full-length TIMP-1 molecule.
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spelling doaj-art-dd2a337b917b44c985698d96bbf78e092025-08-20T03:24:26ZengWileyThe Scientific World Journal2356-61401537-744X2014-01-01201410.1155/2014/467907467907Functional Characterization of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) N- and C-Terminal Domains during Xenopus laevis DevelopmentM. A. Nieuwesteeg0J. A. Willson1M. Cepeda2M. A. Fox3S. Damjanovski4Department of Biology, University of Western Ontario, 1151 Richmond Street, London, ON, N6A 5B7, CanadaDepartment of Biology, University of Western Ontario, 1151 Richmond Street, London, ON, N6A 5B7, CanadaDepartment of Biology, University of Western Ontario, 1151 Richmond Street, London, ON, N6A 5B7, CanadaDepartment of Biology, University of Western Ontario, 1151 Richmond Street, London, ON, N6A 5B7, CanadaDepartment of Biology, University of Western Ontario, 1151 Richmond Street, London, ON, N6A 5B7, CanadaExtracellular matrix (ECM) remodeling is essential for facilitating developmental processes. ECM remodeling, accomplished by matrix metalloproteinases (MMPs), is regulated by endogenous tissue inhibitors of metalloproteinases (TIMPs). While the TIMP N-terminal domain is involved in inhibition of MMP activity, the C-terminal domain exhibits cell-signaling activity, which is TIMP and cell type dependent. We have previously examined the distinct roles of the Xenopus laevis TIMP-2 and -3 C-terminal domains during development and here examined the unique roles of TIMP-1 N- and C-terminal domains in early X. laevis embryos. mRNA microinjection was used to overexpress full-length TIMP-1 or its individual N- or C-terminal domains in embryos. Full-length and C-terminal TIMP-1 resulted in increased lethality compared to N-terminal TIMP-1. Overexpression of C-terminal TIMP-1 resulted in significant decreases in mRNA levels of proteolytic genes including TIMP-2, RECK, MMP-2, and MMP-9, corresponding to decreases in MMP-2 and -9 protein levels, as well as decreased MMP-2 and MMP-9 activities. These trends were not observed with the N-terminus. Our research suggests that the individual domains of TIMP-1 are capable of playing distinct roles in regulating the ECM proteolytic network during development and that the unique functions of these domains are moderated in the endogenous full-length TIMP-1 molecule.http://dx.doi.org/10.1155/2014/467907
spellingShingle M. A. Nieuwesteeg
J. A. Willson
M. Cepeda
M. A. Fox
S. Damjanovski
Functional Characterization of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) N- and C-Terminal Domains during Xenopus laevis Development
The Scientific World Journal
title Functional Characterization of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) N- and C-Terminal Domains during Xenopus laevis Development
title_full Functional Characterization of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) N- and C-Terminal Domains during Xenopus laevis Development
title_fullStr Functional Characterization of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) N- and C-Terminal Domains during Xenopus laevis Development
title_full_unstemmed Functional Characterization of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) N- and C-Terminal Domains during Xenopus laevis Development
title_short Functional Characterization of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) N- and C-Terminal Domains during Xenopus laevis Development
title_sort functional characterization of tissue inhibitor of metalloproteinase 1 timp 1 n and c terminal domains during xenopus laevis development
url http://dx.doi.org/10.1155/2014/467907
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