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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 RNA and protein expression. (A) Wild type and L16 mutant cells were grown in SD medium at 30 °C. Total RNAs were isolated from cells at the log phase (OD600 = 1.5) and subjected to Northern blotting analysis with a radiolabeled pma1+ probe. An ethidium bromide stained gel shows that total RNA was present as a loading control. (B) Wild type and L16 mutant cells were grown as described above, after which cell lysates were prepared. Pma1 protein expression was assessed by Western blotting using anti-Pma1 serum. Tubulin was used as a loading control.
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f0020: Pma1 RNA and protein expression. (A) Wild type and L16 mutant cells were grown in SD medium at 30 °C. Total RNAs were isolated from cells at the log phase (OD600 = 1.5) and subjected to Northern blotting analysis with a radiolabeled pma1+ probe. An ethidium bromide stained gel shows that total RNA was present as a loading control. (B) Wild type and L16 mutant cells were grown as described above, after which cell lysates were prepared. Pma1 protein expression was assessed by Western blotting using anti-Pma1 serum. Tubulin was used as a loading control.

Mentions: To determine the expression profiles associated with pma1+, wild-type and L16 mutant cells were grown in SD medium, after which pma1+ mRNA levels were determined by Northern hybridization. As shown in Fig. 4A, similar pma1+ mRNA levels were expressed in both L16 mutant and wild type cells. Next, we determined the amounts of Pma1 protein by Western blotting under the same growth conditions. As shown in Fig. 4B, similar amounts of Pma1 protein were expressed in both cell types. On the basis of these results, we concluded that there were no differences in the amounts and stabilities of the Pma1 protein expressed in L16 and wild type cells. This indicated that the specific H+-ATPase activity of the Pma1-L16 protein was lower than that of the wild type Pma1 protein.


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 RNA and protein expression. (A) Wild type and L16 mutant cells were grown in SD medium at 30 °C. Total RNAs were isolated from cells at the log phase (OD600 = 1.5) and subjected to Northern blotting analysis with a radiolabeled pma1+ probe. An ethidium bromide stained gel shows that total RNA was present as a loading control. (B) Wild type and L16 mutant cells were grown as described above, after which cell lysates were prepared. Pma1 protein expression was assessed by Western blotting using anti-Pma1 serum. Tubulin was used as a loading control.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4219986&req=5

f0020: Pma1 RNA and protein expression. (A) Wild type and L16 mutant cells were grown in SD medium at 30 °C. Total RNAs were isolated from cells at the log phase (OD600 = 1.5) and subjected to Northern blotting analysis with a radiolabeled pma1+ probe. An ethidium bromide stained gel shows that total RNA was present as a loading control. (B) Wild type and L16 mutant cells were grown as described above, after which cell lysates were prepared. Pma1 protein expression was assessed by Western blotting using anti-Pma1 serum. Tubulin was used as a loading control.
Mentions: To determine the expression profiles associated with pma1+, wild-type and L16 mutant cells were grown in SD medium, after which pma1+ mRNA levels were determined by Northern hybridization. As shown in Fig. 4A, similar pma1+ mRNA levels were expressed in both L16 mutant and wild type cells. Next, we determined the amounts of Pma1 protein by Western blotting under the same growth conditions. As shown in Fig. 4B, similar amounts of Pma1 protein were expressed in both cell types. On the basis of these results, we concluded that there were no differences in the amounts and stabilities of the Pma1 protein expressed in L16 and wild type cells. This indicated that the specific H+-ATPase activity of the Pma1-L16 protein was lower than that of the wild type Pma1 protein.

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.