Domain specific overexpression of TIMP-2 and TIMP-3 reveals MMP-independent functions of TIMPs during<i>Xenopus laevis</i>development

Nieuwesteeg, Michelle A.; Walsh, Logan A.; Fox, Mark A; Damjanovski, Sashko

doi:10.1139/o2012-014

Cited by 7 publications

(7 citation statements)

References 44 publications

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“…Using semiquantitative RT-PCR, we assayed for changes in mRNA levels of the MMP inhibitors (TIMP-2, TIMP-3, and RECK), as well as the MMPs (MMP-2, MMP-9, and MT1-MMP). All of these genes have previously been shown to be important regulators of ECM remodeling during development [8, 9, 12, 34]. mRNA levels were normalized to Ef1 α and compared to control (uninjected) embryos.…”

Section: Resultsmentioning

confidence: 99%

“…At the present time there is comparatively little research characterizing the roles of the N- and C-terminal TIMP domains in vivo , particularly as they pertain to development. Our lab has previously examined the specific functions of the N- and C-terminal domains of TIMP-2 and -3 during X. laevis development and found that the two domains have unique functions when overexpressed individually in X. laevis embryos [12]. To date, there has been no comparison of the unique functions of TIMP-1 N- and C-terminal domains in a developmental context.…”

Section: Discussionmentioning

confidence: 99%

“…As the catalytic domains of MMPs are highly conserved between species, it is not surprising that TIMP-1 N-terminal MMP-inhibitory domains are also well conserved [37]. We have previously shown that the TIMP-2 N-terminal domains are also more highly conserved across species than their C-terminal domains, whereas, for TIMP-3, we found that the C-terminal domains are more highly conserved [12]. Our findings suggest that TIMP-1 may behave more like TIMP-2 than TIMP-3 with respect to MMP inhibition.…”

Section: Discussionmentioning

confidence: 99%

“…Our lab has previously preformed a series of domain specific overexpression experiments, which suggest that TIMP-2 and -3 N- and C-terminal domains may have specific MMP-dependent and independent functions, respectively, during embryogenesis in X. laevis [12]. In the present study, we use a similar approach to characterize the unique functions of the TIMP-1 N- and C-terminal domains during early X. laevis development.…”

Section: Introductionmentioning

confidence: 96%

“…Knockout of membrane-bound MMP-14 (MT1-MMP) during mouse development is embryonic lethal [8–10], as is knockout of RECK due to defects in angiogenesis [11]. Additionally, our lab has shown that overexpression of TIMP-2 and -3 during Xenopus laevis development leads to axial and neural tube defects [12]. Thus, disruption of activity levels in this proteolytic ECM remodeling network is detrimental during development.…”

Section: Introductionmentioning

confidence: 99%

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Functional Characterization of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) N- and C-Terminal Domains duringXenopus laevisDevelopment

Nieuwesteeg

Willson

Cepeda

et al. 2014

The Scientific World Journal

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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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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Functional Characterization of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) N- and C-Terminal Domains duringXenopus laevisDevelopment

Nieuwesteeg

Willson

Cepeda

et al. 2014

The Scientific World Journal

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The Clinical Utility of TIMP3 Expression in ACTH-Secreting Pituitary Tumor

et al. 2015

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In recent years, the tissue inhibitor of metalloproteinase-3 (TIMP3) plays a pivotal role in tumorigenesis, while the role of TIMP3 in adrenocorticotrophic hormone (ACTH)-secreting pituitary adenomas remains unclear. In this study, 86 sporadic pituitary tumor specimens, including ACTH (40), GH (18), PRL-secreting (8), and non-functioining (20) and non-tumorous pituitary samples (n = 10) were available, and then, the mRNA and protein expression of TIMP3 was quantified by quantitative reverse transcriptase polymerase chain reaction (RT-PCR), western blotting, and immunohistochemistry, respectively. Our findings showed that TIMP3 expression was significantly correlated with Ki-67 expression and the invasiveness of pituitary adenomas. TIMP3 mRNA and protein expression were reduced in ACTH-secreting pituitary adenomas and the other three types of pituitary adenomas compared to adjacent non-tumorous pituitary tissues (all p < .01). On the other hand, the expression of TIMP3 was negatively correlated with tumor size and Ki-67 in ACTH-secreting pituitary adenomas. TIMP3 mRNA expression was significantly lower in invasive pituitary adenomas than that in noninvasive ones (1.92-fold, p < .05). TIMP3 protein levels were also significantly lower in the majority of invasive adenomas (1.41-fold, p < .05) Furthermore, TIMP3 mRNA and protein expression were significantly lower in pituitary giant adenomas than those in microadenomas (2.58-fold, p < .05). In conclusion, the expression of TIMP3 is low in pituitary adenomas including ACTH-secreting pituitary adenomas and negatively associated with tumor aggressiveness. TIMPs may play a potential role in the progression of ACTH-secreting pituitary adenomas and be useful as a biomarker of invasiveness.

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Stable expression of α1-antitrypsin Portland in MDA-MB-231 cells increased MT1-MMP and MMP-9 levels, but reduced tumour progression.

Willson

Muir

Evered

et al. 2017

J. Cell Commun. Signal.

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The membrane bound matrix metalloproteinase MT1-MMP plays roles in modulating cell movement, independent of its abilities to remodel the extracellular matrix. Unlike many MMPs, MT1-MMP is activated in the Golgi prior to secretion by a pro-protein convertase, primarily furin. Regulation of the activation of pro-MT1-MMP has been methodically investigated, as altering the level of the active protein has broad implications in both activating other pro-MMPs, including pro-MMP-2, and many subsequent remodelling events. Our previous work in MCF-7 cells has demonstrated that modest, and not extremely high, levels of active MT1-MMP manifests into altered cell morphology and movement. At this low but optimal amount of MT1-MMP protein, changes to MT1-MMP levels are always mirrored by MMP-9 and pERK levels, and always opposite to MMP-2 levels. In this study, stable expression of the furin inhibitor α1-antitrypsin Portland (α1-PDX) in MDA-MB-231 cells increased overall MT1-MMP levels, but cells maintained a 21% proportion of pro-MT1-MMP. The increase in MT1-MMP was mirrored by increases in MMP-9 and pERK, but a decrease in MMP-2. These changes were associated with increased NF-κB transcription. In vitro analysis showed that α1-PDX decreased cell protrusions and migration, and this manifested as decreased tumourigenesis when examined in vivo using a chick CAM assay.

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Domain specific overexpression of TIMP-2 and TIMP-3 reveals MMP-independent functions of TIMPs duringXenopus laevisdevelopment

Cited by 7 publications

References 44 publications

Functional Characterization of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) N- and C-Terminal Domains duringXenopus laevisDevelopment

Functional Characterization of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) N- and C-Terminal Domains duringXenopus laevisDevelopment

The Clinical Utility of TIMP3 Expression in ACTH-Secreting Pituitary Tumor

Stable expression of α1-antitrypsin Portland in MDA-MB-231 cells increased MT1-MMP and MMP-9 levels, but reduced tumour progression.

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