The cortactin-binding domain of WIP is essential for podosome formation and extracellular matrix degradation by murine dendritic cells

TY - JOUR

T1 - The cortactin-binding domain of WIP is essential for podosome formation and extracellular matrix degradation by murine dendritic cells

AU - Bañón-Rodríguez, Inmaculada

AU - Monypenny, James

AU - Ragazzini, Chiara

AU - Franco, Ana

AU - Calle, Yolanda

AU - Jones, Gareth E

AU - Antón, Inés M

N1 - Copyright © 2010 Elsevier GmbH. All rights reserved.

PY - 2010/10/19

Y1 - 2010/10/19

N2 - In immature dendritic cells (DCs) podosomes form and turn over behind the leading edge of migrating cells. The Arp2/3 complex activator Wiskott-Aldrich Syndrome Protein (WASP) localises to the actin core of forming podosomes together with WASP-Interacting Protein (WIP). A second weaker Arp2/3 activator, cortactin, is also found at podosomes where it has been proposed to participate in matrix metalloproteinase (MMP) secretion. We have previously shown that WIP(-/-) DCs are unable to make podosomes. WIP binds to cortactin and in this report we address whether WIP regulates cortactin-mediated MMP activity. Using DCs derived from splenic murine precursors, we found that wild-type cells were able to localise MMPs at podosomes where matrix degradation takes place. In contrast, WIP(-/-) DCs remain able to synthesise MMPs but do not degrade the extracellular matrix. Infection of WIP KO DCs with lentivirus expressing WIP restored both podosome formation and their ability to degrade the extracellular matrix, implicating WIP-induced podosomes as foci of functional MMP location. When WIP KO DCs were infected with a mutant form of WIP lacking the cortactin-binding domain (WIPΔ110-170) DCs were only able to elaborate disorganised podosomes that were unable to support MMP-mediated matrix degradation. Taken together, these results suggest a role for WIP not only in WASP-mediated actin polymerisation and podosome formation, but also in cortactin-mediated extracellular matrix degradation by MMPs.

AB - In immature dendritic cells (DCs) podosomes form and turn over behind the leading edge of migrating cells. The Arp2/3 complex activator Wiskott-Aldrich Syndrome Protein (WASP) localises to the actin core of forming podosomes together with WASP-Interacting Protein (WIP). A second weaker Arp2/3 activator, cortactin, is also found at podosomes where it has been proposed to participate in matrix metalloproteinase (MMP) secretion. We have previously shown that WIP(-/-) DCs are unable to make podosomes. WIP binds to cortactin and in this report we address whether WIP regulates cortactin-mediated MMP activity. Using DCs derived from splenic murine precursors, we found that wild-type cells were able to localise MMPs at podosomes where matrix degradation takes place. In contrast, WIP(-/-) DCs remain able to synthesise MMPs but do not degrade the extracellular matrix. Infection of WIP KO DCs with lentivirus expressing WIP restored both podosome formation and their ability to degrade the extracellular matrix, implicating WIP-induced podosomes as foci of functional MMP location. When WIP KO DCs were infected with a mutant form of WIP lacking the cortactin-binding domain (WIPΔ110-170) DCs were only able to elaborate disorganised podosomes that were unable to support MMP-mediated matrix degradation. Taken together, these results suggest a role for WIP not only in WASP-mediated actin polymerisation and podosome formation, but also in cortactin-mediated extracellular matrix degradation by MMPs.

KW - Animals

KW - Carrier Proteins

KW - Cell Line, Tumor

KW - Cell Surface Extensions

KW - Cortactin

KW - Dendritic Cells

KW - Enzyme Precursors

KW - Extracellular Matrix

KW - Fibronectins

KW - Gelatinases

KW - Humans

KW - Matrix Metalloproteinase 2

KW - Matrix Metalloproteinase 9

KW - Mice

KW - Mice, Knockout

KW - Protein Binding

KW - Protein Structure, Tertiary

U2 - 10.1016/j.ejcb.2010.09.001

DO - 10.1016/j.ejcb.2010.09.001

M3 - Article

C2 - 20952093

SN - 0171-9335

VL - 90

SP - 213

EP - 223

JO - European Journal of Cell Biology

JF - European Journal of Cell Biology

IS - 2-3

ER -