A NUCLEAR MATRIX PROTEIN RELATED TO INTERMEDIATE FILAMENTS PROTEINS IS A MEMBER OF THE COMPLEX BINDING ALPHOID DNAIN VITRO
✍ Scribed by N.I. Enukashvily; I.B. Lobov; A. Kukalev; O.I. Podgornaya
- Publisher
- Elsevier Science
- Year
- 2000
- Tongue
- English
- Weight
- 520 KB
- Volume
- 24
- Category
- Article
- ISSN
- 1065-6995
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✦ Synopsis
Abstract
A complex of three proteins (of 80, 70, 58kDa—p80, p70, and p58, respectively) with the ability to bind alphoid DNA (α‐satDNA) was revealed by gel mobility shift assay (GMSA) in human nuclear matrix. The probes of the α‐satDNA bound in the GMSA with the greatest specificity, but the complex was capable of binding human satellite 3 fragment. According to ion exchange and affinity chromatography, the complex includes two DNA‐binding proteins, p70 and p80, and a non‐DNA‐binding one, p58, which enhances the specificity of binding to the α‐satDNA. GMSA, SDS‐PAGE and immunoblotting showed that the lamins, as well as constitutive centromeric proteins (CENP‐A, CENP‐B, CENP‐C, CENP‐G), were not incorporated into the complex. It was demonstrated by immunoprecipitation assay that p70 and, probably p58, share a common antigen determinant with the rod domain of intermediate filaments (IF) proteins. The results obtained indicate that the nuclear matrix contains at least one IF‐related protein that is able to bind specifically to α‐satDNA in vitro and that this protein is distinct from the lamins.