Chromatographic Separation of DNA Dependent RNA Polymerases and Molecular Properties of RNA Polymerase II from a Leishmania Spp
IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata
View Archive Info| Field | Value | |
| Title |
Chromatographic Separation of DNA Dependent
RNA Polymerases and Molecular Properties of
RNA Polymerase II from a Leishmania Spp
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| Creator |
Sadhukhan, Pranab K
Chakraborty, Asit K Dasgupta, Arindam Majumder, Hemanta K |
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| Subject |
Molecular & Human Genetics
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| Description |
Multiple forms of DNA-dependent RNA polymerases have been isolated and characterized from Leishmania strain UR6
promastigotes. RNA polymerases from this organism fail to resolve into multiple forms by conventional chromatography on DEAE-Sephadex A25, but could be separated by a modification of the method using CM-Sephadex C25. The CM-Sephadex bound enzyme is resistant to a-amanitin even up to a concentration of 250 µg/ml. The activity which flows through CM-Sephadex further resolves into two forms upon chromatography on DEAE-Sephadex A25. These forms are sensitive to a-amanitin to different extent. Enzyme activity in peak I is 50% inhibited by 3 µg/ml and in peak II by 50 µg/ml of the drug respectively. The
enzyme in peak I has been further purified by heparin agarose and fast performance liquid chromatography (FPLC) on MonoQ.
The enzyme has Stoke’s radius of 70Å, a sedimentation coefficient of 17.6S and an f/fo of 1.35. Analysis of ammonium sulfate
and metal ion optima of the enzyme in peak I, relative activities with Mn+2 versus Mg+2 and template specificities gave results
similar to those reported for other type II RNA polymerases in eukaryotes. The MonoQ purified enzyme resolves into 16
polypeptides on denaturing polyacrylamide gel and densitometric analysis suggests that 9 major bands are present in the stoichiometry
expected of RNA polymerase subunits having molecular weights: 154000; 104000; 77000; 64000; 52000; 48000;
46000; 45000 and 39000 respectively.
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| Date |
1997
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| Identifier |
http://www.eprints.iicb.res.in/804/1/MOLECULAR_AND_CELLULAR_BIOCHEMISTRY%2C_171(_1%2D2)%2C_105%2D114[53].pdf
Sadhukhan, Pranab K and Chakraborty, Asit K and Dasgupta, Arindam and Majumder, Hemanta K (1997) Chromatographic Separation of DNA Dependent RNA Polymerases and Molecular Properties of RNA Polymerase II from a Leishmania Spp. Molecular and Cellular Biochemistry, 171 (1-2). pp. 105-114. |
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| Relation |
http://dx.doi.org/10.1023/A:1006875631531
http://www.eprints.iicb.res.in/804/ |
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