Interaction of the Pisum sativum nuclear matrix proteins with SAR DNA.
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 CuSO4 (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 beta 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.
Rzepecki
R
R
Institute of Biochemistry, Wrocøaw University, Poland.
Markiewicz
E
E
Adamiec
R
R
Szopa
J
J
D034961:Antigens, Nuclear; D002467:Cell Nucleus / Q000737:chemistry; D016384:Consensus Sequence; D004247:DNA / Q000302:isolation & purification* / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D016899:Interferon-beta / Q000235:genetics; D015530:Nuclear Matrix / Q000737:chemistry; D009687:Nuclear Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D018532:Pisum sativum / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D016899:Interferon-beta; D004247:DNA; D003850:Deoxyribonuclease I