Differential regulation of a novel variant of the alpha(6) integrin, alpha(6p).
We have reported previously the existence of an M(r) 70,000 form of the alpha(6) integrin called alpha(6p) in a variety of human epithelial cell lines. Four different experimental conditions were used to examine the regulation of alpha(6) and alpha(6p) integrin. The production of the alpha(6) integrin was decreased by 45% using a protein translation inhibitor (2.25 microM puromycin), whereas production of the alpha(6p) variant was unaffected. The alpha(6p) variant was decreased 60% by actin depolymerization (10 microM cytochalasin D) corresponding to a decrease in its surface expression, whereas alpha(6) integrin production was unaffected. The alpha(6p) variant was resistant to endoglycosidase H treatment, whereas the alpha(6) integrin was both sensitive and resistant to endoglycosidase H treatment, indicating retention in the endoplasmic reticulum and processing through the Golgi apparatus. Additionally, digestion by endoglycosidase F demonstrated both alpha(6p) and alpha(6) integrin contained NH(2)-linked glycosylations and both shifted M(r) approximately 10,000 on enzymatic digestion. Finally, inhibition of serine/threonine phosphatases by either calyculin A (15 nM) or okadaic acid (62 microM) did not affect alpha(6p), whereas the production of alpha(6) integrin was decreased by 50%. These data suggest that the production of the alpha(6p) variant is distinct from alpha(6) integrin and may involve a post-translational processing event at the cell surface.
Davis
Tracy L
TL
Department of Radiation Oncology, Arizona Cancer Center, Tucson, Arizona 85724-5024, USA.
Buerger
Friederike
F
Cress
Anne E
AE
D000199:Actins / Q000378:metabolism; D015703:Antigens, CD / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D000954:Antigens, Surface / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism; D006801:Humans; D039503:Integrin alpha6; D019012:Integrin beta1 / Q000378:metabolism; D016023:Integrins / Q000235:genetics / Q000378:metabolism*; D008387:Marine Toxins; D019319:Okadaic Acid / Q000494:pharmacology; D010080:Oxazoles / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D014407:Tumor Cells, Cultured
D000199:Actins; D015703:Antigens, CD; D000954:Antigens, Surface; D039503:Integrin alpha6; D019012:Integrin beta1; D016023:Integrins; D008387:Marine Toxins; D010080:Oxazoles; C465880:alpha 6p integrin; D019319:Okadaic Acid; D015638:Cytochalasin D; C059041:calyculin A; D006026:Glycoside Hydrolases