Cell adhesion to extracellular matrix regulates the life cycle of integrins.
The expression of alpha 5 beta 1 integrin on the surface of fibroblasts requires adhesion to substratum. We have examined the basis for this adhesion-dependent surface expression by comparing the life cycle of integrins in parallel cultures of adherent and nonadherent cells. Results of biosynthetic labeling experiments in NRK fibroblasts showed that the synthesis and biosynthetic processing of the beta 1 integrin subunit proceed in the absence of cell attachment; however, when examining the behavior of preexisting cell surface integrins, we observed that the alpha beta 1 integrins are internalized and degraded when adhesion to substratum is blocked. A kinetic analysis of integrin internalization in cycloheximide-treated NRK cells showed that each of the fibroblast integrins we examined (in both the beta 1 and beta 3 families) are lost from the cell surface after detachment from substratum. Thus, the default integrin life cycle in fibroblasts involves continuous synthesis, processing, transport to the cell surface, and internalization/degradation. Interestingly, studies with NIH-3T3 cells expressing alpha 1 beta 1 integrin showed that the loss of cell-surface alpha 5 beta 1 integrin is blocked by adhesion of cells to dishes coated with type IV collagen (a ligand for alpha 1 beta 1 integrin) as well as fibronectin. Similarly, adhesion of these cells to dishes coated with type IV collagen stabilizes the surface expression of alpha 5 beta 1 as well as alpha 1 beta 1 integrin. We propose that the adhesion of fibroblasts to extracellular matrix protein alters the integrin life cycle and permits retention of these proteins at the cell surface where they can play important roles in transmitting adhesion-dependent signals.
Dalton
S L
SL
Department of Cell Biology and Anatomy, University of Miami School of Medicine, Florida 33101, USA.
Scharf
E
E
Briesewitz
R
R
Marcantonio
E E
EE
Assoian
R K
RK
D016475:3T3 Cells; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D002448:Cell Adhesion; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D005109:Extracellular Matrix / Q000502:physiology*; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology; D006706:Homeostasis; D039564:Integrin alphaV; D019012:Integrin beta1 / Q000378:metabolism; D039661:Integrin beta3; D016023:Integrins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D007668:Kidney; D007700:Kinetics; D051379:Mice; D008954:Models, Biological; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D051381:Rats; D013997:Time Factors
D015703:Antigens, CD; D039564:Integrin alphaV; D019012:Integrin beta1; D039661:Integrin beta3; D016023:Integrins; D010980:Platelet Membrane Glycoproteins; D011500:Protein Synthesis Inhibitors; D003513:Cycloheximide