GENETIC ANALYSIS OF THE HISTIDINE OPERON IN <i>ESCHERICHIA COLI</i> K12

Goldschmidt, Eugene P.; Cater, Margot; Matney, Thomas S.; Butler, Mary Ann; Greene, A. P.

doi:10.1093/genetics/66.2.219

Cited by 62 publications

(6 citation statements)

References 9 publications

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“…The E. coli FB8 strain (wild-type E. coli K12 UTH1038 [27]) and E. coli FB182 (hisF892) strain [23] were used in the present work. E. coli FB182 carries a single nucleotide deletion in position 718 or 719 of the hisF gene, causing a frame shift and the formation of a premature stop codon, resulting in a shorter (243 aa vs. 258 aa of the wild-type E. coli HisF protein) and non-functioning enzyme [24].…”

Section: Bacterial Strainsmentioning

confidence: 99%

“…To the best of our knowledge, very little is known about the reversion of frameshift mutations resulting in amino acid auxotrophy and the correlation with the structure of the protein encoded by the mutated gene, as well as the growth rate in the absence of the amino acid whose biosynthetic pathway has been altered. For this reason, we investigated the different mutations restoring the prototrophy in an auxotrophic E. coli mutant (strain FB182 [23]) harboring the hisF892 single nucleotide deletion [24], causing a frame shift and resulting in the inability of E. coli cells to grow in the absence of histidine. The hisF gene, involved in the histidine biosynthetic pathway, was chosen as the subject of this study since it encodes a protein whose structure has been clearly elucidated in recent years, especially in the thermophilic bacterium Thermotoga maritima.…”

Section: Introductionmentioning

confidence: 99%

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Effect of Non-Lethal Selection on Spontaneous Revertants of Frameshift Mutations: The Escherichia coli hisF Case

et al. 2022

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Microorganisms possess the potential to adapt to fluctuations in environmental parameters, and their evolution is driven by the continuous generation of mutations. The reversion of auxotrophic mutations has been widely studied; however, little is known about the reversion of frameshift mutations resulting in amino acid auxotrophy and on the structure and functioning of the protein encoded by the revertant mutated gene. The aims of this work were to analyze the appearance of reverse mutations over time and under different selective pressures and to investigate revertant enzymes’ three-dimensional structures and their correlation with a different growth ability. Escherichia coli FB182 strain, carrying the hisF892 single nucleotide deletion resulting in histidine auxotrophy, was subjected to different selective pressures, and revertant mutants were isolated and characterized. The obtained results allowed us to identify different indels of different lengths located in different positions in the hisF gene, and relations with the incubation time and the selective pressure applied were observed. Moreover, the structure of the different mutant proteins was consistent with the respective revertant ability to grow in absence of histidine, highlighting a correlation between the mutations and the catalytic activity of the mutated HisF enzyme.

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Section: Bacterial Strainsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Effect of Non-Lethal Selection on Spontaneous Revertants of Frameshift Mutations: The Escherichia coli hisF Case

et al. 2022

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“…The S. cerevisiae strain RH1616 (MATα aro3-2, ura3-52, his7∆::his7 P -lacZ, gcn4-101) (Kuenzler et al, 1993) was used as recipient for a his7complementing cDNA of a galactose inducible A. nidulans expression library. The E. coli strains HfrG6 (λ -, hisA323) (Matney et al, 1964), SB3931 (λ -, hisF860) and UTH1767 (λ -, hisH1767) (Goldschmidt et al, 1970) with stable mutations in hisA, hisF and hisH, respectively, were obtained from the E. coli Genetic Stock Center (Yale University, New Haven, CT) and served as recipients for the cDNA found by the yeast complementation experiment.…”

Section: Methodsmentioning

confidence: 99%

“…The plasmid DNA pME1611 from one of the transformants was retransformed into RH1616 to verify its ability to complement the histidine auxotrophy. Plasmid pME1611 was further analyzed by transforming it into the histidine auxotrophic E. coli strains HfrG6 (Matney et al, 1964), UTH1767 and SB3931 (Goldschmidt et al, 1970) containing mutations in the hisA, hisH and hisF genes, respectively. Although no functional E. coli promoter was present on the plasmid, the transformed DNA was able to complement the mutations in hisH and hisF, encoding a glutamine amidotransferase and a cyclase, respectively, but not in hisA, encoding imidazole-carboxamide isomerase.…”

Section: Isolation and Characterization Of The Hishf Gene Of A Nidula...mentioning

confidence: 99%

Chromatinstruktur und Regulation der Genexpression am Histidin/Adenin-Verzweigungspunkt in Hefe und Aspergillus

Valerius¹

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who supported me whenever I needed their help.I also especially appreciate the interesting talks with Katrin Düvel and Eric Kübler, the cycling weekends with Christoph Springer in the Swiss mountains, and the introduction into the chromatin experiments by Georg Schnappauf during his time in Zürich. I am thankful to Dr. Klaus Adler in Gatersleben for the friendly cooperation and his excellent Aspergillus pictures.The Deutsche Forschungsgemeinschaft, The Fond der Chemischen Industrie, and the Volkswagenstiftung have granted financial support.

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“…UTH 860 (also called SB3931) (Goldschmidt EP et al, 1970;Garrick-Silversmith L et al, 1970;Thoma R et al 1998) [araC14, glnV44(AS), galK2(Oc), LAM-, hisF860(stable), rpsL145(atrR), malT1 (LamR), xylA5, mtl-1]…”

Section: Methodsmentioning

confidence: 99%

Directed evolution of novel properties starting from HisF of <i>Thermotoga maritima</i> as a structural scaffold

Ling¹

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tHisF, imidazole glycerol phosphate synthase from hyperthermophilic organism T. maritima, with a (βα) 8 -barrel structure was used as scaffold for directed evolution of enzymes. The thisF gene libraries were constructed in phagemid vector pCANTAB 5E and in expression vector pKK223-3 by partially randomizing target positions in thisF gene. To evolve novel properties from the thisF library, a three-stage approach was adopted. In the first stage, tHisF variants that were able to complement eHisF activity in E. coli auxotroph strain UT860 were drawn out. 32 amino acids mutations were observed.A common mutation F86L was seen in 91 out of 98 tHisF variants. A proposed hypothesis was the residue Phe86 may be critical for the adaptation of hyperthermophilic enzyme tHisF to mesophilic environment. In the second stage, a tHisF mutant (termed HA03) with N'- [(5'-phosphoribosyl) formimino] -5aminoimidazole-4-carboxamide ribonucleotide isomerase (HisA) activity was isolated from tHisF library by complementing an E. coli auxotroph strain Hfr G6. DNA sequencing of HA03 revealed that it carried ten amino acid exchanges. In the third stage, a transient state analog for target reaction (Aldol condensation) was designed and synthesized for future use in bio-panning to select desired biocatalyst candidates; meanwhile, in attempt to evolve aldolase from tHisF library by genetic selection, dihydrodipicolinate synthase (DHDPS, DapA) was targeted, a ∆dapA E. coli strain lacking DHDPS gene was constructed for this project, the biochemical assay system for DHDPS activity was established including the synthesis of an unstable substrate. ZusammenfassungthisF, der Imidazol-Glyzerol-Phosphat-Synthase aus dem hyperthermophilen Organismus T. maritima mit der Struktur einer (ßα) 8 -Barrels, wurde als Gerüst für eine direkte Evolution von Enzymen genutzt. Die thisF-Genbibliotheken wurden durch teilweises Randomisieren von Zielpositionen im thisF-Gen im Phagemid-Vektor pCANTAB 5E und im Expressionsvektor pKK223-3 konstruiert. Um neue Eigenschaften aus der thisF Bibliothek zu evolvieren, wurde ein dreistufiger Ansatz vorgenommen. Auf der ersten Stufe wurden tHisF-Varianten ausgewählt, die in der Lage waren eHisF-Aktivität im auxotrophen E. coli-Stamm UT860 zu komplementieren. 32 Aminosäure-Austausche wurden beobachtet. Eine weit verbreitete Mutation, F86L, wurde in 91 von 98 Varianten von tHisF beobachtet. Eine mögliche Theorie war, dass der Rest Phe86 entscheidend für die Anpassung des hyperthermophilen Enzyms tHisF an die mesophile Umgebung ist. Im zweiten Schritt wurde eine tHisF Mutante (genannt HA03) mit N'- [(5'-phosphoribosyl) formimino] -5-aminoimidazol-4-carboxamidribonukleotid-isomerase (HisA)-Aktivität aus der HisF-Bibliothek durch Komplementation des auxotrophen E. coli Stammes Hfr G6 isoliert. Die Sequenzierung der DNA von HA03 ergab, dass hierbei zehn Aminosäure-Austausche vorlagen. Im dritten Schritt wurde für zukünftige Anwendungen ein Übergangszustands-Analogon für eine Zielreaktion (Aldo1-Kondensation) konstruiert und synthetisi...

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GENETIC ANALYSIS OF THE HISTIDINE OPERON IN ESCHERICHIA COLI K12

Cited by 62 publications

References 9 publications

Effect of Non-Lethal Selection on Spontaneous Revertants of Frameshift Mutations: The Escherichia coli hisF Case

Effect of Non-Lethal Selection on Spontaneous Revertants of Frameshift Mutations: The Escherichia coli hisF Case

Chromatinstruktur und Regulation der Genexpression am Histidin/Adenin-Verzweigungspunkt in Hefe und Aspergillus

Directed evolution of novel properties starting from HisF of <i>Thermotoga maritima</i> as a structural scaffold

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