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A new pma1 mutation identified in a chronologically long-lived fission yeast mutant.

Naito C, Ito H, Oshiro T, Ohtsuka H, Murakami H, Aiba H - FEBS Open Bio (2014)

Bottom Line: We isolated a chronologically long-lived mutant of Schizosaccharomyces pombe and found a new mutation in pma1 (+) that encoded for an essential P-type proton ATPase.An Asp-138 to Asn mutation resulted in reduced Pma1 activity, concomitant with an increase in the chronological lifespan of this fission yeast.This study corroborates our previous report indicating Pma1 activity is crucial for the determination of life span of fission yeast, and offers information for better understanding of the enzyme, Pma1.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Microbiology, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan.

ABSTRACT
We isolated a chronologically long-lived mutant of Schizosaccharomyces pombe and found a new mutation in pma1 (+) that encoded for an essential P-type proton ATPase. An Asp-138 to Asn mutation resulted in reduced Pma1 activity, concomitant with an increase in the chronological lifespan of this fission yeast. This study corroborates our previous report indicating Pma1 activity is crucial for the determination of life span of fission yeast, and offers information for better understanding of the enzyme, Pma1.

No MeSH data available.


Pma1 activities. (A) Pma1 activities in wild type (open bars) and L16 mutant (filled bars) were assayed using cell lysates prepared from cells grown in SD medium until OD600 = 1 or 2. Results are the means ± s.d.’s of three independent experiments. Statistical analyses were performed using the Student’s t-test and indicated as follows: N.S. nonsignificant; ∗∗P < 0.01. (B) Pma1 activities in wild type (open bars) and L16 mutant (filled bars) were assayed using cell lysates prepared from cells grown in SD medium that contained the indicated glucose concentrations (%). Experiments were repeated twice, with similar results. The representative data are shown.
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f0015: Pma1 activities. (A) Pma1 activities in wild type (open bars) and L16 mutant (filled bars) were assayed using cell lysates prepared from cells grown in SD medium until OD600 = 1 or 2. Results are the means ± s.d.’s of three independent experiments. Statistical analyses were performed using the Student’s t-test and indicated as follows: N.S. nonsignificant; ∗∗P < 0.01. (B) Pma1 activities in wild type (open bars) and L16 mutant (filled bars) were assayed using cell lysates prepared from cells grown in SD medium that contained the indicated glucose concentrations (%). Experiments were repeated twice, with similar results. The representative data are shown.

Mentions: Pma1 is a P-type plasma membrane ATPase that is a hydrogen ion pump [11,12]. H+-ATPase activity of the L16 mutant was analyzed to characterize the effect of the pma1-L16 mutation. Cell lysates were prepared from cells after growth to the mid-logarithmic phase (OD600 = 1) and to the stationary phase (OD600 = 2), after which their H+-ATPase activities were determined. As shown in Fig. 3A, the ATPase activity of the L16 mutant was lower than that of wild type cells at the stationary phase. In S. cerevisiae, it is known that Pma1 H+-ATPase activity is positively regulated by the glucose concentration in the medium; when glucose is added to carbon-starved cells, their ATPase activity increases [13,14]. Thus, we assessed S. pombe Pma1 activity in response to the glucose concentration.


A new pma1 mutation identified in a chronologically long-lived fission yeast mutant.

Naito C, Ito H, Oshiro T, Ohtsuka H, Murakami H, Aiba H - FEBS Open Bio (2014)

Pma1 activities. (A) Pma1 activities in wild type (open bars) and L16 mutant (filled bars) were assayed using cell lysates prepared from cells grown in SD medium until OD600 = 1 or 2. Results are the means ± s.d.’s of three independent experiments. Statistical analyses were performed using the Student’s t-test and indicated as follows: N.S. nonsignificant; ∗∗P < 0.01. (B) Pma1 activities in wild type (open bars) and L16 mutant (filled bars) were assayed using cell lysates prepared from cells grown in SD medium that contained the indicated glucose concentrations (%). Experiments were repeated twice, with similar results. The representative data are shown.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4219986&req=5

f0015: Pma1 activities. (A) Pma1 activities in wild type (open bars) and L16 mutant (filled bars) were assayed using cell lysates prepared from cells grown in SD medium until OD600 = 1 or 2. Results are the means ± s.d.’s of three independent experiments. Statistical analyses were performed using the Student’s t-test and indicated as follows: N.S. nonsignificant; ∗∗P < 0.01. (B) Pma1 activities in wild type (open bars) and L16 mutant (filled bars) were assayed using cell lysates prepared from cells grown in SD medium that contained the indicated glucose concentrations (%). Experiments were repeated twice, with similar results. The representative data are shown.
Mentions: Pma1 is a P-type plasma membrane ATPase that is a hydrogen ion pump [11,12]. H+-ATPase activity of the L16 mutant was analyzed to characterize the effect of the pma1-L16 mutation. Cell lysates were prepared from cells after growth to the mid-logarithmic phase (OD600 = 1) and to the stationary phase (OD600 = 2), after which their H+-ATPase activities were determined. As shown in Fig. 3A, the ATPase activity of the L16 mutant was lower than that of wild type cells at the stationary phase. In S. cerevisiae, it is known that Pma1 H+-ATPase activity is positively regulated by the glucose concentration in the medium; when glucose is added to carbon-starved cells, their ATPase activity increases [13,14]. Thus, we assessed S. pombe Pma1 activity in response to the glucose concentration.

Bottom Line: We isolated a chronologically long-lived mutant of Schizosaccharomyces pombe and found a new mutation in pma1 (+) that encoded for an essential P-type proton ATPase.An Asp-138 to Asn mutation resulted in reduced Pma1 activity, concomitant with an increase in the chronological lifespan of this fission yeast.This study corroborates our previous report indicating Pma1 activity is crucial for the determination of life span of fission yeast, and offers information for better understanding of the enzyme, Pma1.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Microbiology, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan.

ABSTRACT
We isolated a chronologically long-lived mutant of Schizosaccharomyces pombe and found a new mutation in pma1 (+) that encoded for an essential P-type proton ATPase. An Asp-138 to Asn mutation resulted in reduced Pma1 activity, concomitant with an increase in the chronological lifespan of this fission yeast. This study corroborates our previous report indicating Pma1 activity is crucial for the determination of life span of fission yeast, and offers information for better understanding of the enzyme, Pma1.

No MeSH data available.