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Nucleic Acids Research, 1975, Vol. 2, No. 5 655-666
© 1975

Articles

Isolation and characterization of collagen messenger RNA*

L. Wang, C. L. Simoes, S. Sonohara, M. Brentani, H. F. Andrade, Jr., S.M.F. da Silva, J. M. Salles, N Marques** and R. Brentani***

Laboratório de Oncologia Experimental, Faculdade de Medicina, Universidade de São Paulo Caixa Postal 2921, São Paulo, Brasil

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 x 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.

*A preliminary report on some of the present results, has been commincated at the International Symposium on Macromolecules, Rio de Janeiro, July, 1974

**Permanent address: Departamento de Radiologia, Faculdade de Medicine, USP.

***Permanent address: Departamento de Bioquímica, Instituto de Quimica, USP.


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