Isolation and characterization of collagen messenger RNA

Abstract
Chick embryo collagen-synthesizing polysomes were isolated by differential centrifugation. RNA extracted from these particles was chromatographed in oligo(dT)-cellulose columns and the mRNA thus obtained characterized as collagen mRNA by its electrophoretical mobility in acrylamide gels (equivalent to 1.05 × 106 daltons) and its effect upon a cell-free system derived from Krebs ascites tumor cells. The incorporation of 3H-proline was markedly dependent upon rabbit reticulocyte initiation factors and inhibited by initiation inhibitors such as aurintricarboxilate and pyrocatechol violet. The incorporation product was characterized as collagen by its lack of tryptophan, digestibility by purified bacterial collagenase, and by its co-chromatography with unlabeled chick collagen in Sephadex G-200 and CM-cellulose columns.