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The detection and purification of a protein that recognizes a line 1 DNA repetitive sequence

100%

Lukyanov D., Rogolinski J., Podgornaya O., Rzeszowska-Wolny J.

Cellular and Molecular Biology Letters

2000

tom 05

nr 4

551-521

A protein which binds specifically to MspI8 (a 454 bp long repeated sequence highly homologous to the 5’ untranslated region (5'UTR) of the LINE 1 sequence) was found and identified in nuclear extracts of rat liver cells. This protein was detected using the electrophoretic mobility shift assay (EMSA) and was purified by Q-Sepharose and DNA affinity chromatography. Its molecular mass was estimated by SDS electrophoresis as 29 kDa. The possibility that this protein (p29) is the rat analogue of human L1PBP-A, specific for the human LINE sequence LPE1, is discussed.

The nuclear protein p30 specifically interacts with a nuclear matrix attachment region from the rat genome

76%

Fedorov A., Lukyanov D., Rogolinski J., Widlak P., Podgornaya O., Rzeszowska-Wolny J.

Cellular and Molecular Biology Letters

2004

tom 09

nr 1

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.

Ograniczanie wyników

2 Cellular and Molecular Biology Letters

2 Lukyanov D.

2 Podgornaya O.

2 Rogolinski J.

2 Rzeszowska-Wolny J.

1 Fedorov A.

1 Widlak P.

1 2004

1 2000

Wyniki wyszukiwania

The detection and purification of a protein that recognizes a line 1 DNA repetitive sequence

The nuclear protein p30 specifically interacts with a nuclear matrix attachment region from the rat genome