Isolation and characterization of a novel nucleic acid binding protein from calf lenses.

The transitional process of lens cellular differentiation is accompanied by several unique morphological and biochemical changes. Pyknosis or apoptosis of the nucleus involves extensive degradation of genetic materials. In an attempt to search for a gene product responsible for such a regulatory process, we have adopted DNA-cellulose affinity chromatography to enrich the specific binding protein. A binding protein was isolated by high salt (0.8M KCl) wash of the lens polysomal fraction and purified to apparent homogeneity by DNA-cellulose affinity column and chromatofocusing. The nucleic acid binding protein has an apparent molecular weight of 36,000, designated as regulatory factor 36 (RF-36), as determined by SDS/PAGE. Amino acid composition analysis indicated that RF-36 contains high proportions of glycine, alanine, characteristic of the core heteronucleus RNP proteins. Comparative immunological studies with other DNA binding protein antigen (e.g. helix destabilizing protein) suggest the existence of some common overlapping determinant. However, when monoclonal anti-RF-36 was used as immunoprobe, no cross immunoactivity was detected between these homologous binding proteins, suggesting some antigenic diversity among these two nucleic acid binding proteins from different organisms.