Differential synthesis and phosphorylation of the alpha-crystallin A and B chains during bovine lens fiber cell differentiation.
[14C]-amino acids and [32P]-orthophosphate incorporation experiments were carried out in bovine lenses in culture to study the synthesis and phosphorylation of alpha-crystallin A and B polypeptides during differentiation of the lens fiber cells. Following culture, the [14C] or [32P]-labelled alpha-crystallin was isolated by gel filtration chromatography from four regions of the lens corresponding to: A) quiescent epithelial cells, B) dividing epithelial cells and early stages of elongation, C) young elongating fibers, and D) mature fibers from the superficial cortex. The incorporation of label into the alpha-crystallin primary gene products alpha A2 and alpha B2 and their respective phosphorylated forms alpha A1 and alpha B1 was determined by isoelectric focusing and radioautography. Different synthesis and phosphorylation patterns were observed in alpha A and alpha B polypeptides. Synthesis and phosphorylation of the alpha B chain occurs most actively in the epithelial cells, both processes decrease during differentiation and there is no net accumulation of the phosphorylated form alpha B1 in the mature fiber cell. In contrast to the B chain, the A chain synthesis, minimal in the epithelial cell, increases with differentiation. Most striking, the A chain phosphorylation, not detectable in the epithelial cells, increases with differentiation. In the mature fiber cell, the phosphorylated form alpha A1 accounts for one third of the A chain. These observations indicate that the two chains may have different functions. the synthesis and phosphorylation patterns of alpha A suggest a lens-specific function of this polypeptide in the fiber cell and in the terminal differentiation process.(ABSTRACT TRUNCATED AT 250 WORDS)
Chiesa
R
R
Department of Ophthalmology, College of Physicians and Surgeons, Columbia University, New York, NY 10032.
McDermott
M J
MJ
Spector
A
A
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001345:Autoradiography; D002417:Cattle; D002454:Cell Differentiation; D002850:Chromatography, Gel; D003459:Crystallins / Q000096:biosynthesis / Q000378:metabolism*; D007525:Isoelectric Focusing; D007908:Lens, Crystalline / Q000378:metabolism*; D010766:Phosphorylation
D000596:Amino Acids; D003459:Crystallins