Determination of the nucleotide sequence for the glutamate synthase structural genes of Escherichia coli K-12

Guillermo Oliver; Guillermo Gosset; Ray Sanchez-Pescador; Edmundo Lozoya; Lailig M. Ku; Noemi Flores; Baltazar Becerril; Fernando Valle; Francisco Bolivar

doi:10.1016/0378-1119(87)90207-1

Determination of the nucleotide sequence for the glutamate synthase structural genes of Escherichia coli K-12

Guillermo Oliver, Guillermo Gosset, Ray Sanchez-Pescador, Edmundo Lozoya, Lailig M. Ku, Noemi Flores, Baltazar Becerril, Fernando Valle, Francisco Bolivar^*

^*Corresponding author for this work

Medicine, Nephrology Division

Research output: Contribution to journal › Article › peer-review

58 Scopus citations

Abstract

We have determined the complete nucleotide sequence of a 6.3-kb chromosomal HpaI-EcoRI fragment, that contains the structural genes for both the large and small subunits of the Escherichia coli K-12 glutamate synthase (GOGAT) enzyme, as well as the 5 ' - and 3 ' -flanking and intercistronic DNA regions. The M_rs of the two subunits, as deduced from the nucleotide (nt) sequence, were estimated as 166 208 and 52 246. Partial amino acid sequence of the GOGAT enzyme revealed that the large subunit starts with a cysteine residue that is probably generated by a proteolytic cleavage. Northern blotting experiments revealed a transcript of approximately 7300 nt, that at least contains the cistrons for both subunits. A transcriptional start point and a functional promoter were identified in the 5' DNA flanking region of the large subunit gene. The messenger RNA nontranslated leader region has 120 nt and shares identity with the leader regions of E. coli ribosomal operons, in particular around the so-called boxA sequence implicated in antitermination. Other possible regulatory sequences are described.

Original language	English (US)
Pages (from-to)	1-11
Number of pages	11
Journal	Gene
Volume	60
Issue number	1
DOIs	https://doi.org/10.1016/0378-1119(87)90207-1
State	Published - 1987

Keywords

Recombinant DNA
flavoprotein
functional promoter
nitrogen metabolism
oligodeoxynucleotide primer
protein amino terminus
transcriptional start point

ASJC Scopus subject areas

Genetics

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@article{b17b00ef75f04c958e07342f8d23a6f4,

title = "Determination of the nucleotide sequence for the glutamate synthase structural genes of Escherichia coli K-12",

abstract = "We have determined the complete nucleotide sequence of a 6.3-kb chromosomal HpaI-EcoRI fragment, that contains the structural genes for both the large and small subunits of the Escherichia coli K-12 glutamate synthase (GOGAT) enzyme, as well as the 5 ' - and 3 ' -flanking and intercistronic DNA regions. The Mrs of the two subunits, as deduced from the nucleotide (nt) sequence, were estimated as 166 208 and 52 246. Partial amino acid sequence of the GOGAT enzyme revealed that the large subunit starts with a cysteine residue that is probably generated by a proteolytic cleavage. Northern blotting experiments revealed a transcript of approximately 7300 nt, that at least contains the cistrons for both subunits. A transcriptional start point and a functional promoter were identified in the 5' DNA flanking region of the large subunit gene. The messenger RNA nontranslated leader region has 120 nt and shares identity with the leader regions of E. coli ribosomal operons, in particular around the so-called boxA sequence implicated in antitermination. Other possible regulatory sequences are described.",

keywords = "Recombinant DNA, flavoprotein, functional promoter, nitrogen metabolism, oligodeoxynucleotide primer, protein amino terminus, transcriptional start point",

author = "Guillermo Oliver and Guillermo Gosset and Ray Sanchez-Pescador and Edmundo Lozoya and Ku, {Lailig M.} and Noemi Flores and Baltazar Becerril and Fernando Valle and Francisco Bolivar",

note = "Funding Information: We wish to thank Gilbert0 Mosqueda for his help in the purification of the GOGAT enzyme. The protein sequence determination was performed by Guillermo Ramirez in the Protein/DNA Sequence Synthesis Facility of the University of Wisconsin Biotechnology Center, Madison, WI (U.S.A.), supported by funds from the Public Health Service, National Institutes of Health (Shared Equipment grant SlO-RRi684, National Cancer Institute continued support grant CA-07175 and the General Research Support grant to the University of Wisconsin Medical School) and from the University of Wisconsin Graduate School. We wish to thank Dr. Edmund0 Calva who first developed some of the purification manipulations of the GOGAT enzyme at the University of Wisconsin Biotechnology Center. G. Gosset is a recipient of a fellowship from Consejo National de Ciencia y Tecnologia, Mexico. This work was supported in part by Consejo National de Ciencia y Tecnologia, Mexico, Donativo PCCBNAL-022584.",

year = "1987",

doi = "10.1016/0378-1119(87)90207-1",

language = "English (US)",

volume = "60",

pages = "1--11",

journal = "Gene",

issn = "0378-1119",

publisher = "Elsevier",

number = "1",

}

Determination of the nucleotide sequence for the glutamate synthase structural genes of Escherichia coli K-12. / Oliver, Guillermo; Gosset, Guillermo; Sanchez-Pescador, Ray; Lozoya, Edmundo; Ku, Lailig M.; Flores, Noemi; Becerril, Baltazar; Valle, Fernando; Bolivar, Francisco.

In: Gene, Vol. 60, No. 1, 1987, p. 1-11.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Determination of the nucleotide sequence for the glutamate synthase structural genes of Escherichia coli K-12

AU - Oliver, Guillermo

AU - Gosset, Guillermo

AU - Sanchez-Pescador, Ray

AU - Lozoya, Edmundo

AU - Ku, Lailig M.

AU - Flores, Noemi

AU - Becerril, Baltazar

AU - Valle, Fernando

AU - Bolivar, Francisco

N1 - Funding Information: We wish to thank Gilbert0 Mosqueda for his help in the purification of the GOGAT enzyme. The protein sequence determination was performed by Guillermo Ramirez in the Protein/DNA Sequence Synthesis Facility of the University of Wisconsin Biotechnology Center, Madison, WI (U.S.A.), supported by funds from the Public Health Service, National Institutes of Health (Shared Equipment grant SlO-RRi684, National Cancer Institute continued support grant CA-07175 and the General Research Support grant to the University of Wisconsin Medical School) and from the University of Wisconsin Graduate School. We wish to thank Dr. Edmund0 Calva who first developed some of the purification manipulations of the GOGAT enzyme at the University of Wisconsin Biotechnology Center. G. Gosset is a recipient of a fellowship from Consejo National de Ciencia y Tecnologia, Mexico. This work was supported in part by Consejo National de Ciencia y Tecnologia, Mexico, Donativo PCCBNAL-022584.

PY - 1987

Y1 - 1987

N2 - We have determined the complete nucleotide sequence of a 6.3-kb chromosomal HpaI-EcoRI fragment, that contains the structural genes for both the large and small subunits of the Escherichia coli K-12 glutamate synthase (GOGAT) enzyme, as well as the 5 ' - and 3 ' -flanking and intercistronic DNA regions. The Mrs of the two subunits, as deduced from the nucleotide (nt) sequence, were estimated as 166 208 and 52 246. Partial amino acid sequence of the GOGAT enzyme revealed that the large subunit starts with a cysteine residue that is probably generated by a proteolytic cleavage. Northern blotting experiments revealed a transcript of approximately 7300 nt, that at least contains the cistrons for both subunits. A transcriptional start point and a functional promoter were identified in the 5' DNA flanking region of the large subunit gene. The messenger RNA nontranslated leader region has 120 nt and shares identity with the leader regions of E. coli ribosomal operons, in particular around the so-called boxA sequence implicated in antitermination. Other possible regulatory sequences are described.

AB - We have determined the complete nucleotide sequence of a 6.3-kb chromosomal HpaI-EcoRI fragment, that contains the structural genes for both the large and small subunits of the Escherichia coli K-12 glutamate synthase (GOGAT) enzyme, as well as the 5 ' - and 3 ' -flanking and intercistronic DNA regions. The Mrs of the two subunits, as deduced from the nucleotide (nt) sequence, were estimated as 166 208 and 52 246. Partial amino acid sequence of the GOGAT enzyme revealed that the large subunit starts with a cysteine residue that is probably generated by a proteolytic cleavage. Northern blotting experiments revealed a transcript of approximately 7300 nt, that at least contains the cistrons for both subunits. A transcriptional start point and a functional promoter were identified in the 5' DNA flanking region of the large subunit gene. The messenger RNA nontranslated leader region has 120 nt and shares identity with the leader regions of E. coli ribosomal operons, in particular around the so-called boxA sequence implicated in antitermination. Other possible regulatory sequences are described.

KW - Recombinant DNA

KW - flavoprotein

KW - functional promoter

KW - nitrogen metabolism

KW - oligodeoxynucleotide primer

KW - protein amino terminus

KW - transcriptional start point

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U2 - 10.1016/0378-1119(87)90207-1

DO - 10.1016/0378-1119(87)90207-1

M3 - Article

C2 - 3326786

AN - SCOPUS:0023551349

VL - 60

SP - 1

EP - 11

JO - Gene

JF - Gene

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