The adaptor protein fish associates with members of the ADAMs family and localizes to podosomes of Src-transformed cells

Clare L. Abram, Darren F. Seals, Ian Pass, Daniel Salinsky, Lisa Maurer, Therese M. Roth, Sara Courtneidge

Research output: Contribution to journalArticle

166 Citations (Scopus)

Abstract

Fish is a scaffolding protein and Src substrate. It contains an amino-terminal Phox homology (PX) domain and five Src homology 3 (SH3) domains, as well as multiple motifs for binding both SH2 and SH3 domain-containing proteins. We have determined that the PX domain of Fish binds 3-phosphorylated phosphatidylinositols (including phosphatidylinositol 3-phosphate and phosphatidylinositol 3,4-bisphosphate). Consistent with this, a fusion protein of green fluorescent protein and the Fish PX domain localized to punctate structures similar to endosomes in normal fibroblasts. However, the full-length Fish protein was largely cytoplasmic, suggesting that its PX domain may not be able to make intermolecular interactions in unstimulated cells. In Src-transformed cells, we observed a dramatic re-localization of some Fish molecules to actin-rich structures called podosomes; the PX domain was both necessary and sufficient to effect this translocation. We used a phage display screen with the fifth SH3 domain of Fish and isolated ADAM19 as a binding partner. Subsequent analyses in mammalian cells demonstrated that Fish interacts with several members of the ADAMs family, including ADAMs 12, 15, and 19. In Src-transformed cells, ADAM12 co-localized with Fish in podosomes. Because members of the ADAMs family have been implicated in growth factor processing, as well as cell adhesion and motility, Fish could be acting as an adaptor molecule that allows Src to impinge on these processes.

Original languageEnglish (US)
Pages (from-to)16844-16851
Number of pages8
JournalJournal of Biological Chemistry
Volume278
Issue number19
DOIs
StatePublished - May 9 2003
Externally publishedYes

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Fish Proteins
Fish
Fishes
src Homology Domains
Proteins
Endosomes
Phosphatidylinositols
Green Fluorescent Proteins
Cell Adhesion
Bacteriophages
Cell Movement
Actins
Intercellular Signaling Peptides and Proteins
Fibroblasts
Molecules
4-ethoxymethylene-2-phenyl-2-oxazoline-5-one
Podosomes
Cell adhesion
Fusion reactions
Display devices

ASJC Scopus subject areas

  • Biochemistry

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The adaptor protein fish associates with members of the ADAMs family and localizes to podosomes of Src-transformed cells. / Abram, Clare L.; Seals, Darren F.; Pass, Ian; Salinsky, Daniel; Maurer, Lisa; Roth, Therese M.; Courtneidge, Sara.

In: Journal of Biological Chemistry, Vol. 278, No. 19, 09.05.2003, p. 16844-16851.

Research output: Contribution to journalArticle

Abram, Clare L. ; Seals, Darren F. ; Pass, Ian ; Salinsky, Daniel ; Maurer, Lisa ; Roth, Therese M. ; Courtneidge, Sara. / The adaptor protein fish associates with members of the ADAMs family and localizes to podosomes of Src-transformed cells. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 19. pp. 16844-16851.
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