Dimethyl sulfoxide targets phage RNA polymerases to promote transcription.
Dimethyl sulfoxide (DMSO) is a "universal" solvent in pharmaceutical sciences and cell biology. DMSO was previously reported to facilitate in vitro transcription of chromatin and supercoiled plasmid, with the underlying mechanism being attributed to the alteration of the template structure. Here, we demonstrated that low concentrations of DMSO significantly increase the phage polymerase-catalyzed RNA synthesis when the naked and short PCR products were used as templates, suggesting that DMSO promotes transcription through additional mechanism(s). Interestingly, SP6 RNA polymerase was more sensitive to the DMSO stimulation than T7 RNA polymerase, suggesting that the polymerase is an important target for DMSO stimulation of RNA synthesis. Consistent with this finding, we also showed that DMSO dramatically elevated the RNA polymerase activity. This elevated activity is explained by altered polymerase structure as reflected by a change in intrinsic fluorescence. Furthermore, DMSO was shown to simultaneously accumulate both the abortive and full-length transcripts, leading us to conclude that the DMSO-altered polymerase structure primarily encodes an enhanced activity at the stage of transcription initiation. DMSO-induced alteration of the polymerase structure and function highlights a potentially generalized mechanism in interpreting the molecular action of this popular solvent.
State Key Laboratory of Virology and Department of Biotechnology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, China.
Zhiqiang
Z
Chen
Yi
Y
Zhang
D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000187:drug effects* / Q000502:physiology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D014158:Transcription, Genetic / Q000502:physiology; D014764:Viral Proteins / Q000737:chemistry / Q000187:drug effects* / Q000502:physiology
D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D004121:Dimethyl Sulfoxide