B.V. All rights reacrvcd.
Nucleotide (Dihydroorotic DNA)
gene of Saccharomyces
sequence of the URAl acid dehydrogenase
Anne Roy C.h’.R. S.
L’. P. R. Y(103 Cu~hrog~r~Psr
b! J.-P. Lccocq
Mol&~laire er Stnrctu~ule. Irtsritur de Bidogie Molhlrrire
I I December
6 71184 Strmhour~y Cede.\,
1992: RcceiLed at publishers:
The complete nucleotide sequence of the URAl gene encoding dihydroorotic acid dehydrogcnase (DHOdehase) is presented. This enzyme catalyses the conversion of DHO to erotic acid and plays a major role in the pyrimidine pathway. as DHO is the effector of the positive control of the transcription of at least four genes, URAl, URA3, URA4 and URAfO. Comparisons between the amino acid sequence of the yeast DHOdehase and sequences of DHOdehases previously isolated from Dict~wtellum discoi’dum, Escherichia cnli and Bacillus suhtilis reveal no obvious homologies.
The URAI gene of Saccharom)ces cerevisiue encodes the dihydroorotic acid dehydrogenase (DHOdehase), which catalyses the conversion of dihydroorotic acid to erotic acid in the pyrimidine pathway (Lacroute, 1968). URAl expression is regulated at the transcriptional level through a positive control which requires both PPRl (pyrimidine pathway regulator l), the product of the regulatory PPRl gene (Loison et al., 1981; Losson and Lacroutc, 1983) and the inducer DHO (Lacroute, 1968). The induced and noninduced tsp were previously located both in the S. cereviske
Correspondertc~e to: Dr. A. Roy.
hourg Cedex, France. *On rcqucst
C.P.R. Canc&gCn&e et Mutag&& I.B.M.C., 15, Rue Descartes 67084 Stras-
the author will supply dctailcd
Fax (33)-(88).61-06-80. cxpcrimcntal
conclusions reached in this Brief Note. ’ This paper IS dcdicatcd to the mernorq of Dr. J.-P. Lecocq. Ahhrcviations:
aa, amino acid(s);
bp, base pair(s):
dihydroorotic or 1000 bp: nt.
nucleotide( s); ORF. open reading frame; PPR I, pyrimidine pathway rcgulator I: fv/J, transcription start point(a): C,dS,,,,,. upstrcnm activating scqucncc
background and in Schi~osac,charonz),ce.r pomhe (Losson et al., 1985). We sequenced a 2884-bp DNA fragment carrying this gene (Loison et al., 1981) by the dideoxy chain termination method (Sanger et al., 1977). Upstream (205-2 18 bp and 234-249 bp) from the tsp are protein. two binding sites (UAS,,,,, ) of a PPRl-dependent These sites, identified both by correlation to the consensus sequence and by in vitro DNase I footprinting with yeastcell extracts overproducing or not PPRl (Roy et al., 1990), are located in a 2X5-bp DNA fragment upstream from a HirrdIII site (Fig. 1) and are necessary for PPRl induction as shown by DHOdehase activity (data not shown), contrary to the previous results of Loison et al. (1981). An ORF encoding the complete 3 14-aa sequence of the dihydroorotic acid dehydrogenase is found downstream from these tsp. No special features have been noticed in the aa sequence. The hydrophobicity computed from the aa scquence according to the coefficients of Kyte and Doolittle (1982) reveals no unusual pattern (data not shown). The codon bias index calculated as suggested by Bennetzcn and Hall (1982) has a value of -0.013. Comparisons between the S. cerevisiae DHOdehase aa sequence and either the D. discoi’dum or E. coli aa sequences previously reported
150 120 240 360 CG
CAGFTWZX&TA!XAG%TIMlXWZJA~~~ G VAG M S I D EN LN L LR K TM;CTTATGACTIXACIVXCkWiCQ&FC~~~CAAAAAACCI AYD
(Larsen and Jensen. 1985), or a B. suhtiliscluster of p~‘r genes (EMBL data library) reveal no overall significant homologies.
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E‘.w/wrid~itr cc,/; and characterization
Snecial thanks arc due to F. Lacroute for his constant interest and to B. Winsor for her English editing of the manuscript. This work benefited from technical advice from F. Exinger for DHOdehase activity assay.
480 -22 600 -62 720 -102 840 .I42 960 -182
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