Three ATP1 Genes Are Present on Chromosome II in Saccharomyces cerevisiae

Ono

2005

By means of gene disruption analyses, Saccharomyces cerevisiae strain YPH499 was shown to have two, and only two, copies of ATP3 that encodes the γ -subunit of H + -dependent ATP synthase and locates on the right arm of chromosome II. Linkage analyses of the two distinguishably marked copies of ATP3 indicated that the distance between them was about 43 cM. Since YBR030W, an ORF proximal to ATP3 by a distance of 17 kbp, was also found to be duplicated, we marked them with two distinguishable nutritional markers, which were also distinguishable from those used for marking the two copies of ATP3, and achieved four-point linkage analyses; CEN2 marked with an appropriate nutritional marker gene was included as a reference point. And, the following linkage map was deduced: CEN2-[11 cM]-YBR030Wa-[8 cM]-ATP3a-[47 cM]-ATP3b-[55 cM]-YBR030Wb. From this map, we suspected that a segment spanning at least YBR030W-ATP3 would be inversely duplicated on the right arm of chromosome II. We then carried out chromosome fragmentation analyses, using several laboratory strains including YPH499, and obtained data in accord with our speculation for all strains, although the distance between the two copies of ATP3 varied from 48 kbp to 192 kbp among the strains examined.

Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Inverted repeat of a large segment unveiled on the right arm of Saccharomyces cerevisiae chromosome II

Ono

2005

“…The Southern hybridization procedure, the preparation of DIG-labelled ATP3 (2.9 kb XbaI-HindIII fragment), and other probes used in the experiments were carried out as detailed previously (Takeda et al, 1995).…”

Section: Miscellaneousmentioning

confidence: 99%

“…These subunits are encoded ATP1, ATP2, ATP3, ATP16 and ATP15, respectively (Cox et al, 1992). Although they are recognized as being single-copy genes by the yeast genome project, our intensive studies have shown that ATP1 (Takeda et al, 1995(Takeda et al, , 1999 and ATP2 (Takeda M, Katayama H, Satoh T, Oku S, Mabuchi T. Gene organization of ATP2 gene encoding F 1 -ATPase β subunit of Saccharomyces cerevisiae; in preparation) are actually triplicate genes. These findings are significant in two respects: first, there may be a correlation between the subunit number and the gene dosage; and second, the copy number of a given gene reported by the genome project is a matter for reconsideration.…”

Section: Introductionmentioning

confidence: 99%

Studies on the ATP3 gene of Saccharomyces cerevisiae: presence of two closely linked copies, ATP3a and ATP3b, on the right arm of chromosome II

Ishibashi

Kunihiro

et al. 2003

Self Cite

In this paper, we present evidence that there are two closely linked copies of the ATP3 gene coding for the γ subunit of the F 1 F 0 -ATPase complex (EC3.6.1.34) in four laboratory strains of Saccharomyces cerevisiae, even though the yeast genome project has reported that ATP3 is a single-copy gene on chromosome II. We previously reported that the gene dosage (three copies) of ATP1 and ATP2 is coincident with the subunit number of F 1 -α and F 1 -β, but that the gene dosage of ATP3 was not consistent with the subunit stoichiometry of F 1 F 0 -ATPase. By applying long PCR and gene walking analyses, we estimated that the two copies of ATP3 were approximately 20 kb apart, and we designated that which is proximal to the centromere ATP3a, while we named that which is distal ATP3b. The nucleotide sequences of the two copies of ATP3 were identical to the reported sequence in the W303-1A, W303-1B and LL20 strains, while only the DC5 strain had a single base substitution in its ATP3a. With the exception of this substitution, the other nucleotide sequences were identical to the upstream 860 bp and the downstream 150 bp. The differences between ATP3 with the single base substitution (Ser 308 to Phe) and ATP3 without the substitution on the complementation of the ATP3 disruptant and on the maintenance of the mitochondrial DNA were observed, suggesting that Atp3ap and Atp3bp in the DC5 strain might have different functions. However, it should not always be necessary for yeast cells to carry different types of ATP3 because the other three strains carry the same type of ATP3. It was also demonstrated that the disruption of the ATP3 genes basically leads to a loss of wild-type mtDNA, but the stability of the mtDNA is not dependent on the ATP3 alone.

The three copies of theATP1 gene are arranged in tandem on chromosome II ofSaccharomyces cerevisiae S288C

Takeda

Sato

et al. 1999

Self Cite

In the yeast Saccharomyces cerevisiae there are three copies of the F1F0‐ATPase α‐subunit gene ATP1 on chromosome II (Takeda et al., 1995). However, after genome analysis using S. cerevisiae strain S288C, only one ATP1 gene sequence was observed (Feldman et al., 1994; Obermaier et al., 1995). To check whether the number of copies of ATP1 is strain‐dependent or not, we carried out three different experiments: (a) long‐PCR analyses of total DNAs isolated from several reference strains, carried out by preparing 29‐mer oligonucleotides based on the 5′‐ and 3′‐ up‐ and downstream regions of the ATP1 nucleotide sequence using the data from the genome project to synthesize primers; (b) restriction analyses of chromosome II from the reference strains with SplI; and (c) long‐PCR analyses of prime clones 70113 and 70804, both of which contained two ATP1 gene copies, ATP1a and ATP1b, and ATP1b and ATP1c, respectively, using 30 nucleotides just inside the 3′‐end (sense) and 5′‐end (antisense) of the ATP1‐coding region as primers. In the case of the long‐PCR experiments, the reference strains DC5, SEY2102, W303‐1A, W303‐1B, LL20 and DBY746, as well as strain S288C, generated a DNA fragment of approximately 32 kb, which hybridized with ATP1. During SplI digestion, a DNA fragment of more than 50 kb which hybridized with ATP1, was obtained from all reference strains. In the case of prime clone analyses using the long‐PCR experiments, the distance between ATP1a and ATP1b or ATP1b and ATP1c was approximately 10 kb or 7 kb, respepectively. The S288C strain generated these two DNA fragments, as do the other strains. These results showed that all these strains contained three copies of ATP1 on chromosome II. Copyright © 1999 John Wiley & Sons, Ltd.