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PMID: 7690620 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Ligand-dependent and -independent integrin focal contact localization: the role of the alpha chain cytoplasmic domain.

Molecular biology of the cell ·Vol. 4 ·No. 6 ·1993-06-00 ·Pages 593-604

Briesewitz R, Kern A, Marcantonio EE

Abstract

Many integrin receptors localize to focal contact sites upon binding their ligand. However, unoccupied integrin receptors do not localize to focal contact sites. Because the integrin beta 1 cytoplasmic domain appears to have a focal contact localization signal, there must be a mechanism by which this domain is kept inactive in the unoccupied state and becomes exposed or activated in the occupied receptor. We considered that this mechanism involves the alpha subunit cytoplasmic domain. To test this hypothesis, we have established two NIH 3T3 cell lines that express either the human alpha 1 wild-type subunit (HA1 cells) or the cytoplasmic domain deleted alpha 1 subunit (CYT cells). Both cell lines express similar levels of the human alpha 1 subunit, and there is no significant effect of the deletion on the dimerization and surface expression of the receptor. Furthermore, the deletion had no effect on the binding or adhesion via alpha 1 beta 1 to its ligand collagen IV. However, when these two cell lines are plated on fibronectin (FN), which is a ligand for alpha 5 beta 1 but not for alpha 1 beta 1, there is a striking difference in the cellular localization of alpha 1 beta 1. The HA1 cells show only alpha 5 in focal contacts, without alpha 1, demonstrating that all of the integrin localization is ligand dependent. In contrast, when the CYT cells are plated on FN, the mutant alpha 1 appears in focal contacts along with the alpha 5/beta 1. Thus, there is both ligand-dependent (alpha 5/beta 1) and ligand-independent (alpha 1/beta 1) focal contact localization in these cells. The truncated alpha 1 also localized to focal contacts in a ligand-independent manner on vitronectin. We conclude that the mutant alpha 1 no longer requires ligand occupancy for focal contact localization. These data strongly suggest that the alpha cytoplasmic domain plays a role in the normal ligand-dependent integrin focal contact localization.

MeSH Terms
3T3 Cells Amino Acid Sequence Animals Binding Sites Cell Adhesion Chromatography, Affinity Collagen/metabolism Cytoplasm/metabolism Fibronectins/metabolism Glycoproteins/metabolism Humans Integrins/chemistry,genetics,metabolism Laminin/metabolism Mice Microscopy, Fluorescence Molecular Sequence Data Mutagenesis, Site-Directed Vitronectin
Chemicals
Fibronectins Glycoproteins Integrins Laminin Vitronectin Collagen
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Briesewitz R
Department of Pathology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
Kern A
Marcantonio E E
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.4.6.593
Published
1993-06-00
Pages
593-604
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC300966
Subset
IM
Grants
NIGMS NIH HHS · GM 44585 · United States
External Links
PubMed source DOI: 10.1091/mbc.4.6.593 PMC Full Text View journal details
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