Targeting p35/Cdk5 Signalling via CIP-Peptide Promotes Angiogenesis in Hypoxia

Bosutti, A, Qi, J, Pennucci, R, Bolton, D, Matou, S, Ali, K, Tsai, LH, Krupinski, J, Petcu, EB, Montaner, J, Al Baradie, R, Caccuri, F, Caruso, A, Alessandri, G, Kumar, S, Rodriguez, C, Martinez-Gonzalez, J and Slevin, M (2013) Targeting p35/Cdk5 Signalling via CIP-Peptide Promotes Angiogenesis in Hypoxia. PLoS ONE, 8 (9). ISSN 1932-6203

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Official URL: https://e-space.mmu.ac.uk/617493/1/Targeting%20p35...

Abstract

Cyclin-dependent kinase-5 (Cdk5) is over-expressed in both neurons and microvessels in hypoxic regions of stroke tissue and has a significant pathological role following hyper-phosphorylation leading to calpain-induced cell death. Here, we have identified a critical role of Cdk5 in cytoskeleton/focal dynamics, wherein its activator, p35, redistributes along actin microfilaments of spreading cells co-localising with p(Tyr15)Cdk5, talin/integrin beta-1 at the lamellipodia in polarising cells. Cdk5 inhibition (roscovitine) resulted in actin-cytoskeleton disorganisation, prevention of protein co-localization and inhibition of movement. Cells expressing Cdk5 (D144N) kinase mutant, were unable to spread, migrate and form tube-like structures or sprouts, while Cdk5 wild-type over-expression showed enhanced motility and angiogenesis in vitro, which was maintained during hypoxia. Gene microarray studies demonstrated myocyte enhancer factor (MEF2C) as a substrate for Cdk5-mediated angiogenesis in vitro. MEF2C showed nuclear co-immunoprecipitation with Cdk5 and almost complete inhibition of differentiation and sprout formation following siRNA knock-down. In hypoxia, insertion of Cdk5/p25-inhibitory peptide (CIP) vector preserved and enhanced in vitro angiogenesis. These results demonstrate the existence of critical and complementary signalling pathways through Cdk5 and p35, and through which coordination is a required factor for successful angiogenesis in sustained hypoxic condition. © 2013 Bosutti et al.

Item Type:	Article
Peer-reviewed:	Yes
Date Deposited:	06 Dec 2016 09:03
Publisher:	Public Library of Science
Additional Information:	This is an Open Access article published in PloS One, published by Public Library of Science, copyright The Author(s).
Divisions:	Faculties > Science and Engineering
Subject terms:	Science & Technology, Multidisciplinary Sciences, Science & Technology - Other Topics, MULTIDISCIPLINARY SCIENCES, CYCLIN-DEPENDENT KINASE-5, MYOCYTE ENHANCER FACTOR-2, ACUTE ISCHEMIC-STROKE, TRANSCRIPTION FACTOR, ENDOTHELIAL-CELLS, ACTIVATOR P35, IN-VITRO, BRAIN, CDK5, EXPRESSION, Cell Line, Pseudopodia, Cytoskeleton, Humans, Anoxia, Purines, Peptides, Adaptor Proteins, Signal Transducing, Cell Cycle Proteins, DNA Primers, Microscopy, Fluorescence, Blotting, Western, Microarray Analysis, Colorimetry, Analysis of Variance, Immunoprecipitation, Signal Transduction, Cell Movement, RNA Interference, Neovascularization, Physiologic, Mutation, Missense, Cyclin-Dependent Kinase 5, Stroke, Actin Cytoskeleton, MEF2 Transcription Factors, Actin Cytoskeleton, Adaptor Proteins, Signal Transducing, Analysis of Variance, Anoxia, Blotting, Western, Cell Cycle Proteins, Cell Line, Cell Movement, Colorimetry, Cyclin-Dependent Kinase 5, Cytoskeleton, DNA Primers, Humans, Immunoprecipitation, MEF2 Transcription Factors, Microarray Analysis, Microscopy, Fluorescence, Mutation, Missense, Neovascularization, Physiologic, Peptides, Pseudopodia, Purines, RNA Interference, Signal Transduction, Stroke, General Science & Technology, MD Multidisciplinary
URI:	https://e-space.mmu.ac.uk/id/eprint/617493
DOI:	https://doi.org/10.1371/journal.pone.0075538
ISSN	1932-6203

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