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

100%

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.

In vitro DNA binding of purified CcpA protein from Lactococcus lactis IL1403

59%

Kowalczyk M., Borcz B., Plochocka D., Bardowski J.

Acta Biochimica Polonica

2007

tom 54

nr 1

During this study His-tagged CcpA protein purified under native conditions to obtain a biologically active protein was used for molecular analysis of CcpA-dependent regulation. Using electrophoretic mobility shift assays it was demonstrated that CcpA of L. lactis can bind DNA in the absence of the HPr-Ser-P corepressor and exhibits DNA-binding affinity for nucleotide sequences lacking cre sites. However, purified HPr-Ser-P protein from Bacillus subtilis was shown to slightly increase the DNA-binding capacity of the CcpA protein. It was also observed that CcpA bound to the cre box forms an apparently more stable complex than that resulting from unspecific binding. Competition gel retardation assay performed on DNA sequences from two PEP:PTS regions demonstrated that the ybhE, bglS, rheB, yebE, ptcB and yecA genes situated in these regions are most probably directly regulated by CcpA.

Ograniczanie wyników

1 Acta Biochimica Polonica

1 Cellular and Molecular Biology Letters

1 Bardowski J.

1 Borcz B.

1 Fedorov A.

1 Kowalczyk M.

1 Lukyanov D.

1 Plochocka D.

1 Podgornaya O.

1 Rogolinski J.

1 Rzeszowska-Wolny J.

1 Widlak P.

1 2007

1 2004

Wyniki wyszukiwania

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

In vitro DNA binding of purified CcpA protein from Lactococcus lactis IL1403