The nuclear protein p30 specifically interacts with a nuclear matrix attachment region from the rat genome.
In our previous study, a 454 bp DNA fragment was isolated from rat genomic DNA as an element which interacts with nuclear matrix proteins, i.e. a Matrix Associated Region (MAR). Computer analyses revealed that the right half of this fragment, named RME (Rat MAR Element), possesses a high matrix association potential and is likely to be responsible for the matrix association of the whole sequence. RME was used as a probe in an electrophoretic mobility shift assay (EMSA), and with the use of Southwestern blotting, a rat liver nuclear protein which binds specifically to it was identified. Its molecular mass was estimated by SDS-PAGE as 30 kDa (p30). Polyclonal antibodies raised against protein-RME complexes caused a super-shift of specific complexes in EMSA, and bound to p30 in nuclear extracts of rat liver in Western blotting. The immunofluorescence labelling of a rat embryonic fibroblast cell monolayer with anti-p30 antibody revealed a mainly intranuclear pattern of staining.
Fedorov
Anton
A
Institute of Cytology, Russian Academy of Science, Tikhoretsky pr. 4, 194064 St. Petersburg, Russia. a_slon@pochtamt.ru
Lukyanov
Dmitri
D
Rogoliński
Jacek
J
Widłak
Piotr
P
Podgornaya
Olga
O
Rzeszowska-Wolny
Joanna
J
D000818:Animals; D002467:Cell Nucleus / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005347:Fibroblasts / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D045170:Matrix Attachment Regions / Q000235:genetics*; D015530:Nuclear Matrix / Q000235:genetics* / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D051381:Rats
D002843:Chromatin; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins