Nucleotide sequence of the Bacillus subtilis argC gene encoding N-acetylglutamate-gamma-semialdehyde dehydrogenase


The B. subtilis argC gene encoding N-acetylglutamate-gammasemialdehyde dehydrogenase is required for the third step in the biosynthesis of arginine. argC lies within the argCAEBD-cpa-argF cluster at approximately 100 degrees on the chromosome and is expressed from an arginine regulated promoter located just upstream of argC (1,2,3). The argC gene was originally cloned by complementation of an argC~ arginine auxotroph of E. coli with a 1474 bp insert in pBR322 to form pUL710 (1). Tn5 insertions in pUL710 abolished the complementation by the cloned gene and localised the argC gene on the insert DNA (1). Analysis of plasmid encoded products in E. coli minicells using pUL710 and Tn5 insertions indicated the synthesis of a 41 kd protein (M.CM.Smith, A.Mountain, S.Baumberg, unpublished observations). The cloned insert in pUL710 has been completely sequenced using the Sanger chain termination method on both strands of overlapping Sau3A, TaqI and Hpall clones in M13 (4). The sequence presented here represents the only ORF of the predicted size encoded by the insert. The derived amino acid sequence has 39% identity and 62.5% similarity to the E. coli argC sequence (5). The use of the assigned initiation codon, TTG, has been discussed previously (3). The predicted amino acid sequence encoded between 535 and 561 bp (underlined) is completely conserved between the E. coli and the B. subtilis sequences and is similar to 'Box A' of the nucleotide binding pocket consensus sequence (6). 
1 TTGAAAATAGGAATTGTAGGTGCTACAGGATATGGAGGCACCGAACTTGTCAGGATTCTT AACTTTTATCCTTAACATCCACGATGTCCTATACCTCCGTGGCTTGAACAGTCCTAAGAA M K I G I V G A T G Y G G T E L V R I L 61 TCGCATCATCCTCATGCAGAGGAATGCATACTTTATTCATCCAGCGGAGAAGGGAATGTC AGCGTAGTAGGAGTACGTCTCCTTACGTATGAAATAAGTAGGTCGCCTCTTCCCTTACAG S H H P H A E E C I L Y S S S G E G N V 121 TATAGCGAGGGTTATCCTCATCTTACCGGCTTAGCGGATCAGCAGCTGAAGCCGATTGAT ATATCGCTCCCAATAGGAGTAGAATGGCCGAATCGCCTAGTCGTCGACTTCGGCTAACTA Y S E G Y P H L T G L A D Q Q L K P I D 181 ATGAAT ACGATC AAAC ACGAAATAGATATCATGTTTCTCGCTGCGCCGCCCGGAGTATC A TACTTATGCTAGTTTGTGCTTTATCTATAGTACAAAGAGCGACGCGGCGGGCCTCATAGT M N T I K H E I D I M F L A A P P G V S 241 AGTGAATTGACTCCAAAGCTGGCAGACGCGGGAATTACGGTTATTGATCTGTCAGGTGAT TCACTTAACTGAGGTTTCGACCGTCTGCGCCCTTAATGCCAATAACTAGACAGTCCACTA S E L T P K L A D A G I T V I D L S G D 301 CTGAGGATAAAAGAACCGGCTGAATATGAAAAATGGTATAAACGGACAGCGGCACCGAAG GACTCCTATTTTCTTGGCCGACTTATACTTTTTACCATATTTGCCTGTCGCCGTGGCTTC L R I K E P A E Y E K W Y K R T A A P K 3 61 GCGGTGATTCAAGAGGCGGTATACGGTCTGGCAGAACTGAATCAACTGCAAATTCAACAG CGCCACTAAGTTCTCCGCCATATGCCAGACCGTCTTGACTTAGTTGACGTTTAAGTTGTC A V I Q E A V Y G L A E L N Q L Q I Q Q 421 GCGAAACTCATTGCCAATCCAGGCTGTTTTCCAACAGCTGTTTTGCTTGGCCTCGCGCCA CGCTTTGAGTAACGGTTAGGTCCGACAAAAGGTTGTCGACAAAACGAACCGGAGCGCGGT A K L I A N P G C F P T A V L L G L A P 4 81 TTGGCTCAAAAGAAACTGCTCGATGAATCTTTCGTTATCGTTGACGCGAAGACCGGTGTT AACCGAGTTTTCTTTGACGAGCTACTTAGAAAGCAATAGCAACTGCGCTTCTGGCCACAA L A Q K K L L D E S F V I V D A K T G V 5 41 TCCGGAGCGGGAAGAAAAGCATCCATGGGAACTCATTTTTCTGAGCTGAACGACAATTTT AGGCCTCGCCCTTCTTTTCGTAGGTACCCTTGAGTAAAAAGACTCGACTTGCTGTTAAAA S G A G R K A S M G T H F S E L N D N F 601 AAAATTTATAAAGTCAATGAACATCAGCACACGCCGGAAATTGAGCAGGCGCTGAATGAA TTTTAAATATTTCAGTTACTTGTAGTCGTGTGCGGCCTTTAACTCGTCCGCGACTTACTT K I Y K V N E H Q H T P E I E Q A L N E 561 TGGCAGCCAGGTCTCGGGCCCATTACATTTTCGGCTCACTTGTTTCCGATGACAAGGGGC ACCGTCGGTCCAGAGCCCGGGTAATGTAAAAGCCGAGTGAACAAAGGCTACTGTTCCCCG W Q P G L G P I T F S A H L F P M T R G 721 ATCATGGCGACGATGTATACCAGATTAACCTGTGACCTAACCGCAGATGACCTGCATGAT TAGTACCGCTGCTACATATGGTCTAATTGGACACTGGATTGGCGTCTACTGGACGTACTA I M A T M Y T R L T C D L T A D D L H D 781 TTATATTCGG AATTTTACC AAGATTCATATTTTGTGAGAGTGAGGCCAAAAGGTC AGT AC AATATAAGCCTTAAAATGGTTCTAAGTATAAAACACTCTCACTCCGGTTTTCCAGTCATG L Y S E F Y Q D S Y F V R V R P K G Q Y i 41 CCGCAAACGAAAGAAGTGTACGGCAGCAATTTCTGTGATATCGCCGTGACCCTCGATGAG GGCGTTTGCTTTCTTCACATGCCGTCGTTAAAGACACTATAGCGGCACTGGGAGCTACTC P Q T K E V Y G S N F C D I A V T L D E )01 AGAACGAACAGAGTCACGATCGTCTCGGTAATCGATAATTTAATGAAGGGTGCCGCCGGT TCTTGCTTGTCTCAGTGCTAGCAGAGCCATTAGCTATTAAATTACTTCCCACGGCGGCCA R T N R V T I V S V I D N L M K G A A G ) 61 C AGGCAGTGCAAAACTTTAATTTGATGAATGGCTGGAATGAAGAAACTGGACTCACCATC GTCCGTCACGTTTTGAAATTAAACTACTTACCGACCTTACTTCTTTGACCTGAGTGGTAG Q A V Q N F N L M N G W N E E T G L T I )

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  To whom correspondence should be addressed at the Department of Biological and Molecular Science, University of Stirling, Stirling FK9 4LA, UK + Present address: Celltech Group pic, 216 Bath Road, Slough SL1 4EN, UK Downloaded from https://academic.oup.com/nar/article/18/15/4595/1049021 by Indiana University -Bloomington user on 30 August 2023 


ACKNOWLEDGEMENTS

This work was supported by MRC grants G82/0499/8CB to SB and G84/0925/0CB to SB and AM.
Margaret C M Smith, Andrew Mountain, Simon Baumberg
Nucleotide sequence of the Bacillus subtilis argC gene encoding N-acetylglutamate-gamma-semialdehyde dehydrogenase