Enzymatic properties of a proteolytically nicked RNA polymerase of bacteriophage T7.

Ikeda, Richard A.; Richardson, Charles C.

doi:10.1016/s0021-9258(18)61425-5

Cited by 90 publications

(47 citation statements)

References 32 publications

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“…The overall rate constants received compare to an average T7 RNAP progression of around 60 s −1 , as reported for transcription of T7 DNA (Ikeda and Richardson, 1987). A rough estimate for the rate constant of chain growth using T7 RNAP was found to be 100 nucleotides per second (Chamberlin and Ring, 1973).…”

Section: Maximum Transcription Ratesupporting

confidence: 53%

“…Although apparent affinity constants for substrate nucleotides have been estimated for T7 RNAP in previous studies (Chamberlin and Ring, 1973;Ikeda and Richardson, 1987), the therein applied conditions suggest some caution in the interpretation of reported results. One restriction associated with these studies arises from the use of initial substrate concentrations that may have been insufficient to guarantee substrate saturation.…”

Section: Parametermentioning

confidence: 89%

“…One notable exception to the literature data is the affinity constant for GTP, which was estimated to be 76 M from our experiments. In contrast, a raised K M value, ranging from 160 to 190 M, has been typically found for GTP when using bacteriophage T7 RNA polymerase (Chamberlin and Ring, 1973;Ikeda and Richardson, 1987). The higher halfsaturation constant was commonly attributed to a particular involvement of GTP in transcript initiation.…”

Section: Parametermentioning

confidence: 98%

“…Furthermore, T7 RNAP has been applied routinely in cell-free gene expression systems for the production of sensitive protein products, as reviewed earlier (Jermutus et al, 1998;Stiege and Erdmann, 1995). Among the issues examined, particular focus has been placed on promoter recognition as part of the initiation process (Basu and Maitra, 1986;Ikeda and Richardson, 1987;Jia and Patel, 1997;Martin and Coleman, 1987;Maslak and Martin, 1994;Sen and Dasgupta, 1993) and the occurrence of abortive transcription products (Sousa et al, 1992), which are short oligonucleotides synthesized up to 8 to 14 nucleotides in length and released before entering a processive elongation mode. GTP is the initiator nucleotide that is incorporated in the growing RNA chain (Basu and Maitra, 1986;Jia and Patel, 1997;Martin and Coleman, 1987;Sen and Dasgupta, 1993).…”

Section: Introductionmentioning

confidence: 99%

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Kinetic modeling and simulation of in vitro transcription by phage T7 RNA polymerase

Arnold

Siemann

Scharnweber

et al. 2001

Biotechnol. Bioeng.

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This study provides a mathematical model of T7 RNA polymerase (T7 RNAP) kinetics under in vitro conditions targeted at application of this model to simulation of dynamic transcription performance. A functional dependence of transcript synthesis rate is derived based on: (a) essential reactant concentrations, including T7 RNAP and its promoter, substrate nucleotides, and the inhibitory byproduct inorganic pyrophosphate; (b) a distinction among vector characteristics such as recognition sequences regulating transcription initiation and termination, respectively; and (c) specific properties of the nucleotide sequence including both transcript length and nucleotide composition. Inactivation kinetics showed a half‐life of T7 RNAP activity of 50 min under the conditions applied in vitro using the isolated enzyme. Model parameters and their precision are estimated using dynamic simulation and nonlinear regression analysis. The particular novelty of this model is its capability to incorporate linear genomic sequence information for simulation of nonlinear in vitro transcription kinetics. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 72: 548–561, 2001.

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Section: Maximum Transcription Ratesupporting

confidence: 53%

Section: Parametermentioning

confidence: 89%

Section: Parametermentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

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Kinetic modeling and simulation of in vitro transcription by phage T7 RNA polymerase

Arnold

Siemann

Scharnweber

et al. 2001

Biotechnol. Bioeng.

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“…Sam Rabkin showed that plasmids containing a T7 replication origin could be replicated by the T7 replication proteins in vivo, providing a way to identify the secondary origins that function when the primary origin is deleted (120,121). During this period, Rick Ikeda used a proteolytically nicked RNA polymerase to examine its interaction with its promoter (122)(123)(124).…”

Section: Onset Of Studies On Bacteriophage T7 Dna Replicationmentioning

confidence: 99%

Untitled

2015

Annu. Rev. Biochem.

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I spent my childhood and adolescence in North and South Carolina, attended Duke University, and then entered Duke Medical School. One year in the laboratory of George Schwert in the biochemistry department kindled my interest in biochemistry. After one year of residency on the medical service of Duke Hospital, chaired by Eugene Stead, I joined the group of Arthur Kornberg at Stanford Medical School as a postdoctoral fellow. Two years later I accepted a faculty position at Harvard Medical School, where I remain today. During these 50 years, together with an outstanding group of students, postdoctoral fellows, and collaborators, I have pursued studies on DNA replication. I have experienced the excitement of discovering a number of important enzymes in DNA replication that, in turn, triggered an interest in the dynamics of a replisome. My associations with industry have been stimulating and fostered new friendships. I could not have chosen a better career.

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Evolution of C‐Terminal Modification Tolerance in Full‐Length and Split T7 RNA Polymerase Biosensors

Disare

Dickinson

2019

ChemBioChem

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show abstract

Enzymatic properties of a proteolytically nicked RNA polymerase of bacteriophage T7.

Cited by 90 publications

References 32 publications

Kinetic modeling and simulation of in vitro transcription by phage T7 RNA polymerase

Kinetic modeling and simulation of in vitro transcription by phage T7 RNA polymerase

Untitled

Evolution of C‐Terminal Modification Tolerance in Full‐Length and Split T7 RNA Polymerase Biosensors

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