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Archaeal histone stability, DNA binding, and transcription inhibition above 90 degrees C.
Extremophiles. 1998 May; 2(2):75-81.E

Abstract

The DNA binding and compacting activities of the recombinant (r) archaeal histones rHMfA and rHMfB from Methanothermus fervidus, and rHPyA1 from Pyrococcus species GB-3a, synthesized in Escherichia coli, have been shown to be completely resistant to incubation for 4h at 95 degrees C in the presence of 1M KCl. Continued incubation of rHMfA and rHMfB at 95 degrees C resulted in a gradual loss of these activities, and rHMfA and rHMfB lost activity more rapidly at 95 degrees C when the salt environment was reduced to 200 mM K Cl. rHPyA1, in contrast, retained full activity even after a 60-h incubation at 95 degrees C in 1 M KCl, and reducing the salt concentration did not affect the heat resistance of rHPyA1. rHPya1-DNA complexes remained intact at 100 degrees C, and rHPyA1 bound to the template DNA in in vitro transcription reaction mixtures assembled using Pyrococcus furiosus components at 90 degrees C. Transcription in vitro from the P. furiosus gdh promoter was reduced by rHPyA1 binding, in a manner that was dependent on the histone-to-DNA ratio and on the topology of the DNA template. Transcription from circular templates was more sensitive to rHPyA1 binding than transcription from a linear template, consistent with rHPyA1 binding introducing physical barriers to transcription and causing changes in the topology of circular templates that also reduced transcription.

Authors+Show Affiliations

Department of Microbiology, Ohio State University, Columbus 43210, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

9672681

Citation

Soares, D, et al. "Archaeal Histone Stability, DNA Binding, and Transcription Inhibition Above 90 Degrees C." Extremophiles : Life Under Extreme Conditions, vol. 2, no. 2, 1998, pp. 75-81.
Soares D, Dahlke I, Li WT, et al. Archaeal histone stability, DNA binding, and transcription inhibition above 90 degrees C. Extremophiles. 1998;2(2):75-81.
Soares, D., Dahlke, I., Li, W. T., Sandman, K., Hethke, C., Thomm, M., & Reeve, J. N. (1998). Archaeal histone stability, DNA binding, and transcription inhibition above 90 degrees C. Extremophiles : Life Under Extreme Conditions, 2(2), 75-81.
Soares D, et al. Archaeal Histone Stability, DNA Binding, and Transcription Inhibition Above 90 Degrees C. Extremophiles. 1998;2(2):75-81. PubMed PMID: 9672681.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Archaeal histone stability, DNA binding, and transcription inhibition above 90 degrees C. AU - Soares,D, AU - Dahlke,I, AU - Li,W T, AU - Sandman,K, AU - Hethke,C, AU - Thomm,M, AU - Reeve,J N, PY - 1998/7/22/pubmed PY - 1998/7/22/medline PY - 1998/7/22/entrez SP - 75 EP - 81 JF - Extremophiles : life under extreme conditions JO - Extremophiles VL - 2 IS - 2 N2 - The DNA binding and compacting activities of the recombinant (r) archaeal histones rHMfA and rHMfB from Methanothermus fervidus, and rHPyA1 from Pyrococcus species GB-3a, synthesized in Escherichia coli, have been shown to be completely resistant to incubation for 4h at 95 degrees C in the presence of 1M KCl. Continued incubation of rHMfA and rHMfB at 95 degrees C resulted in a gradual loss of these activities, and rHMfA and rHMfB lost activity more rapidly at 95 degrees C when the salt environment was reduced to 200 mM K Cl. rHPyA1, in contrast, retained full activity even after a 60-h incubation at 95 degrees C in 1 M KCl, and reducing the salt concentration did not affect the heat resistance of rHPyA1. rHPya1-DNA complexes remained intact at 100 degrees C, and rHPyA1 bound to the template DNA in in vitro transcription reaction mixtures assembled using Pyrococcus furiosus components at 90 degrees C. Transcription in vitro from the P. furiosus gdh promoter was reduced by rHPyA1 binding, in a manner that was dependent on the histone-to-DNA ratio and on the topology of the DNA template. Transcription from circular templates was more sensitive to rHPyA1 binding than transcription from a linear template, consistent with rHPyA1 binding introducing physical barriers to transcription and causing changes in the topology of circular templates that also reduced transcription. SN - 1431-0651 UR - https://www.unboundmedicine.com/medline/citation/9672681/Archaeal_histone_stability_DNA_binding_and_transcription_inhibition_above_90_degrees_C_ DB - PRIME DP - Unbound Medicine ER -