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Isolation of RNA polymerases from the water mold Achlya
1Botanisches Institut der Universität, Meckenheimer Allee 170, D-5300, Bonn 1, Federal Republic of Germany.
Abstract:
The DNA-dependent RNA polymerases I, II, and III (ribonucleosidetriphosphate: RNA nucleotidyl-transferase, EC 2.7.7.6) from Achlya ambisexualis E87 (male), have been isolated. The highly purified RNA polymerase I was found to be composed of polypeptides with the following molecular weights (·10(-4)): 18.5, 14, 11.8, 7.3, 6.1, 4.9, 4.4, 2.8. RNA polymerase II showed a 400-fold higher resistance against α-amanitin than mammalian or higher plant RNA polymerase II.
Insights
Researchers isolated DNA-dependent RNA polymerases I, II, and III from Achlya ambisexualis. RNA polymerase II from this fungus exhibited significantly higher resistance to α-amanitin compared to mammalian and plant counterparts.
Area of Science:
- Molecular Biology
- Mycology
- Biochemistry
Background:
- DNA-dependent RNA polymerases are crucial enzymes for gene transcription in eukaryotes.
- Understanding the structure and properties of these enzymes across different species can reveal evolutionary insights and functional differences.
Purpose of the Study:
- To isolate and characterize DNA-dependent RNA polymerases I, II, and III from the fungus Achlya ambisexualis E87 (male).
- To investigate the biochemical properties of these isolated enzymes, particularly their sensitivity to α-amanitin.
Main Methods:
- Isolation of DNA-dependent RNA polymerases I, II, and III from Achlya ambisexualis.
- Purification of RNA polymerase I and determination of its polypeptide composition via molecular weight analysis.
- Assessment of RNA polymerase II resistance to α-amanitin in comparison to mammalian and plant RNA polymerases.
Main Results:
- Successfully isolated DNA-dependent RNA polymerases I, II, and III from Achlya ambisexualis.
- The purified RNA polymerase I comprises polypeptides with specific molecular weights (18.5, 14, 11.8, 7.3, 6.1, 4.9, 4.4, 2.8 x 10(-4)).
- RNA polymerase II from Achlya ambisexualis demonstrated a 400-fold greater resistance to α-amanitin than its mammalian or higher plant counterparts.
Conclusions:
- The study successfully characterized key RNA polymerases from Achlya ambisexualis.
- The distinct α-amanitin resistance of Achlya ambisexualis RNA polymerase II suggests unique structural or functional adaptations compared to other eukaryotes.
