Integrin alpha6 cleavage: a novel modification to modulate cell migration.
Integrins play a major role in cell adhesion and migration. Previous work reported that a cleaved form of integrin alpha6 (alpha6p) was detected in invasive human prostate cancer tissue, absent in normal prostate tissue and was produced by urokinase-type Plasminogen Activator (uPA) in a plasmin-independent manner. Using site-directed mutagenesis we identified amino acid residues R594 and R595, located in the "stalk" region of integrin alpha6, as essential for cleavage. The cleavage site is located on the extracellular region of the protein between the beta-barrel domain and the thigh domain. Prostate cancer cells (PC3N) were stably transfected to overexpress the cleavable, wild-type (PC3N-alpha6-WT) or the non-cleavable form of integrin alpha6 (PC3N-alpha6-RR). The number of cells invading laminin 111- and laminin 332-coated filters by PC3N-alpha6-WT cells increased by threefold as compared to PC3N-alpha6-RR cells. Plasminogen activator inhibitor-1 (PAI-1) reduced the invasion of PC3N-alpha6-WT cells by approximately 42% through laminin 332-coated filters and plasmin inhibitor aprotinin had no significant effect. Linear cell migration increased production of integrin alpha6p in the PC3N-alpha6-WT cells and not in the PC3N-alpha6-RR cells and 32% of the PC3N-alpha6-WT cells migrated on laminin 111 in the linear migration assay as compared to the 5% PC3N-alpha6-RR cells. These data taken together suggest that the uPA-mediated cell surface cleavage of the alpha6 integrin extracellular domain is involved in tumor cell invasion and migration on laminin.
Pawar
Sangita C
SC
Dept. of Cell Biology and Anatomy, University of Arizona, Tucson, AZ 85724, USA.
Demetriou
Manolis C
MC
Nagle
Raymond B
RB
Bowden
G Tim
GT
Cress
Anne E
AE
D000595:Amino Acid Sequence; D000596:Amino Acids; D002460:Cell Line; D002465:Cell Movement; D006801:Humans; D039503:Integrin alpha6 / Q000235:genetics / Q000378:metabolism*; D020014:K562 Cells; D008297:Male; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D010641:Phenotype; D010960:Plasminogen Activators / Q000494:pharmacology*; D011471:Prostatic Neoplasms / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid
D000596:Amino Acids; D039503:Integrin alpha6; D010960:Plasminogen Activators