Abstract
GMP synthetase (EC 6.3.4.1), a glutamine amido-transferase encoded by the guaA gene, catalyzes the synthesis of GMP from XMP. The guaA gene was subcloned from the Clarke and Carbon (Clarke, L., and Carbon, J. (1976) Cell 9, 91-99) plasmid pLC34-10, and the nucleotide sequence was determined. The structural gene encodes a protein of 525 amino acid residues having a calculated Mr of 58,604. The amino acid sequence of the NH2 terminus of GMP synthetase was determined and used to verify the translation start site determined from the DNA sequence. A 68-base pair intercistronic region separates guaA from the upstream guaB gene in the polycistronic guaBA operon. The 3' end of the guaA mRNA was determined by S1 nuclease mapping. The 3' end of guaA mRNA is 36-37 nucleotides downstream of the translation stop codon within a region of dyad symmetry that resembles a rho-independent transcription termination site.
Highlights
From the $Departmentof Biochemistry and Molecular Biology, LouisianaState University Schoolof Medicine, Shreveport, Louisiana 71130 and the llDepartmentof Biochemistry, Purdue University, West Lafayette, Indiana 47907
Transferase encoded by the guaA gene, catalyzes the coliguaA structural gene
The desubcloned from the Clarke and Carbon (ClaLrk.,ea,nd duced amino acid sequence contains 525 residues having a Carbon, J. (1976) Cell 9, 91-99) plasmid pLC34-10, andthenucleotidesequence was determinedT. he structural gene encodes a protein of 525 amino acid residues having a calculatedM, of 58,604
Summary
O M AGC TGC CCG TTT OEC ATT A M i GCT M C W A GAA AM Coc TTC TAT GCC GTA CAG TTC HIS PRO GLU VAL THR H I S THR ARC GLM'GLY MET ARC MET L I U GLU ARC PHI VAL 110 ASP CAC CCG GAA GTG ACT CAT ACC CGC CAG 001 A T G CGC A T G CTG GAG COT U T GTG COT GAT. 679 I L I CY8 GLN CY8 CLU ALA LEU TUP T H R PRO ALA LY8 XLI I L I ASP ASP ALA VAL ALA ARC. 1x.I ARC GLU GLII VAL GLr ASP A#P LXI VAL I L I LEU GLY LIU EIR oLr GLK VAL ASP sau ATC Eu: GAG CAG GTA GCC GAC GAT M A oic &TC CTC GCC CTC TCT ffiT OGT Oto GAT TCC. 799 S I R VAL T H R ALA MET LEU LIUHIS ARC ALA I L I GLY LYS ASY LEU THRCIS VAL P H I VAL TCC GTA Ur Q A A C CTG CTG CAC Coc GCT A T C GGT M A M C CTG ACT TUC GTA TTC 0%
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