Multiple factors contribute to integrin-talin interactions in vivo.
The cytoplasmic protein talin is an essential part of the integrin-cytoskeleton link. We characterized the interaction between integrin and two conserved regions of talin, the N-terminal ;head' domain and the C-terminus, which includes the I/LWEQ domain, within the living organism. Green-fluorescent-protein-tagged head and C-terminal domains were recruited to integrin adhesion sites. Both required integrins for recruitment, but the C-terminal domain also required endogenous talin, showing it was not recruited directly by integrins. We used chimeric transmembrane proteins containing the cytoplasmic domain of the integrin beta subunit to examine the integrin-talin head interaction. Monomeric chimeric proteins did not recruit talin head, whereas dimeric chimeras efficiently recruited it and caused a strong inhibition of integrin-mediated adhesion. These chimeras recruited surprisingly few integrin-associated proteins, indicating that recruitment of talin did not initiate a cascade of recruitment. Mutagenesis of the integrin cytoplasmic domain, within the chimera, showed the dominant-negative inhibition was not due to talin sequestration alone and that additional interactions are required.
The Wellcome Trust/Cancer Research UK Gurdon Institute and Department of Anatomy, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QN, UK.
Guy
G
Tanentzapf
Maria D
MD
Martin-Bermudo
Marcus S
MS
Hicks
Nicholas H
NH
Brown
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism*; D004330:Drosophila; D049452:Green Fluorescent Proteins / Q000378:metabolism; D016023:Integrins / Q000235:genetics / Q000378:metabolism*; D047668:Inteins; D008969:Molecular Sequence Data; D009119:Muscle Contraction; D009132:Muscles / Q000196:embryology / Q000502:physiology*; D009154:Mutation; D011485:Protein Binding; D016608:Talin / Q000235:genetics / Q000378:metabolism*
D015815:Cell Adhesion Molecules; D016023:Integrins; D016608:Talin; D049452:Green Fluorescent Proteins