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Modification of the lipid component modulates nuclear matrix nucleolytic activity

100%

Lubocka J., Szopa J., Kozubek A.

Acta Biochimica Polonica

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1995

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tom 42

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nr 2

It was shown that lipid composition of plant nuclear matrix depends on procedure of its isolation. The matrix isolated with the use of lithium diodosalicylate (LiS) differs in its lipid composition from the preparation isolated with the use of nonionic detergent (Triton X-100). It was also shown that the nucleolytic activity of the matrix is related to its lipid component. Matrix depleted in lipids loses half of its nucleolytic activity which is recovered after supplementation with previously extracted lipids. The extent of recovery of the nucleolytic activity is also dependent on the presence of residual DNA in matrix preparation. The recoveries of nucleolytic activities were higher in matrices not depleted in their DNA content.

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Interaction of the Pisum sativum nuclear matrix proteins with SAR DNA

100%

Rzepecki R., Markiewicz E., Adamiec R., Szopa J.

Acta Biochimica Polonica

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1995

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tom 42

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nr 1

We have isolated the nuclear matrices from Pisum sativum cell nuclei using three methods: i. standard procedure involving extraction of cell nuclei with 2 M NaCl and 1% Triton X-100; ii. the same with pretreatment of cell nuclei with 0.5 mM CuS04 (stabilisation step); and iii. method including lithium diiodosalicylate extraction. We compared the polypeptide pattern and residual DNA content of the nuclear matrices isolated. The nuclear matrices displayed a specific endonuclease activity which was due to the presence of a 32 kDa protein. The isolated nuclear matrices bound specifically the scaffold-attached (SAR) DNA derived from human p interferon gene, in the exogenous SAR binding assay. Using the DNA-protein binding blot assay we demonstrated the presence of two nuclear matrix proteins of 66 kDa and 62 kDa which bound specifically SAR DNA.

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Identification of the proteins responsible for SAR DNA binding in nuclear matrix of Cucurbita pepo

80%

Rzepecki R., Markiewicz E., Szopa J.

Acta Biochimica Polonica

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1995

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tom 42

|

nr 2

The nuclear matrices from White bush (Cucurbita pepovar. patisonina)cell nuclei have been isolated using three methods: I, standard procedure involving extraction of cell nuclei with 2 NaCl and 1% X-100; II, the same with pre-treatment of cell nuclei with 0.5 CuS04 (stabilisation step); and III, method with extraction by lithium diiodosalicylate (LIS), and compared the polypeptide pattern. The isolated matrices specifically bind SAR DNA from human β-interferon gene in the exogenous SAR assay and in the gel mobility shift assay. Using IgG against the 32 endonuclease we have found in the DNA-protein blot assay that this protein is one of the proteins binding SAR DNA. have identified three proteins with molecular mass of 65 , 60 and 32 which are responsible for SAR DNA in the gel mobility shift assay experiments.

Ograniczanie wyników

3 Acta Biochimica Polonica

3 Szopa J.

2 Markiewicz E.

2 Rzepecki R.

1 Adamiec R.

1 Kozubek A.

1 Lubocka J.

3 1995

Modification of the lipid component modulates nuclear matrix nucleolytic activity

Interaction of the Pisum sativum nuclear matrix proteins with SAR DNA

Identification of the proteins responsible for SAR DNA binding in nuclear matrix of Cucurbita pepo