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RSC Chromatin-Remodeling Complex Is Important for Mitochondrial Function in Saccharomyces cerevisiae.

Imamura Y, Yu F, Nakamura M, Chihara Y, Okane K, Sato M, Kanai M, Hamada R, Ueno M, Yukawa M, Tsuchiya E - PLoS ONE (2015)

Bottom Line: RSC (Remodel the Structure of Chromatin) is an ATP-dependent chromatin remodeling complex essential for the growth of Saccharomyces cerevisiae.The rsc2Δ phenotypes were relieved by RSC1 overexpression, indicating that the isoforms play a redundant role in respiratory growth.Genome-wide expression analysis in nps1-13 under respiratory conditions suggested that RSC regulates the transcription of some target genes of the HAP complex, a transcriptional activator of respiratory gene expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima, 739-8530, Japan.

ABSTRACT
RSC (Remodel the Structure of Chromatin) is an ATP-dependent chromatin remodeling complex essential for the growth of Saccharomyces cerevisiae. RSC exists as two distinct isoforms that share core subunits including the ATPase subunit Nps1/Sth1 but contain either Rsc1or Rsc2. Using the synthetic genetic array (SGA) of the non-essential mutation method, we screened for mutations exhibiting synthetic growth defects in combination with the temperature-sensitive mutant, nps1-105, and found connections between mitochondrial function and RSC. rsc mutants, including rsc1Δ, rsc2Δ, and nps1-13, another temperature-sensitive nps1 mutant, exhibited defective respiratory growth; in addition, rsc2Δ and nps1-13 contained aggregated mitochondria. The rsc2Δ phenotypes were relieved by RSC1 overexpression, indicating that the isoforms play a redundant role in respiratory growth. Genome-wide expression analysis in nps1-13 under respiratory conditions suggested that RSC regulates the transcription of some target genes of the HAP complex, a transcriptional activator of respiratory gene expression. Nps1 physically interacted with Hap4, the transcriptional activator moiety of the HAP complex, and overexpression of HAP4 alleviated respiratory defects in nps1-13, suggesting that RSC plays pivotal roles in mitochondrial gene expression and shares a set of target genes with the HAP complex.

No MeSH data available.


Related in: MedlinePlus

Overexpression of HAP4 alleviated the respiratory defect of nps1-13.(A) Effect of HAP4 overexpression on nps1-13 growth on YPEG. WT (BY4743) and nps1-13 (BYI-3) cells harboring pRS426 (WT/v and nps1-13/v, respectively) or pRS426GPDpr::HAP4 (WT/HAP4 and nps1-13/HAP4, respectively) were grown to log phase in SD-Ura medium, spotted on YPD and YPEG plates in serial five-fold dilutions and incubated at 30°C for 3 days. (B) Effect of HAP4 overexpression on petit nps1-13 colony formation. The strains described in (A) were plated on YPEG; three independent colonies were subsequently picked, pre-cultured in SD-Ura medium, and separately grown in YPD medium. On the indicated days, 200 cells from each culture were plated on YPD plates and incubated at 30°C for 3 days. Data are presented as the means ± SEM (n = 3).
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pone.0130397.g005: Overexpression of HAP4 alleviated the respiratory defect of nps1-13.(A) Effect of HAP4 overexpression on nps1-13 growth on YPEG. WT (BY4743) and nps1-13 (BYI-3) cells harboring pRS426 (WT/v and nps1-13/v, respectively) or pRS426GPDpr::HAP4 (WT/HAP4 and nps1-13/HAP4, respectively) were grown to log phase in SD-Ura medium, spotted on YPD and YPEG plates in serial five-fold dilutions and incubated at 30°C for 3 days. (B) Effect of HAP4 overexpression on petit nps1-13 colony formation. The strains described in (A) were plated on YPEG; three independent colonies were subsequently picked, pre-cultured in SD-Ura medium, and separately grown in YPD medium. On the indicated days, 200 cells from each culture were plated on YPD plates and incubated at 30°C for 3 days. Data are presented as the means ± SEM (n = 3).

Mentions: To understand the relationship between the RSC and HAP complex, we first determined whether the hap4Δ mutant exhibits similar phenotypes as those induced by mitochondrial dysfunction in rsc mutants. The hap4Δ mutant also exhibited growth defects on a medium containing a non-fermentable carbon source and a high frequency of mtDNA loss (S1 Fig). Next, we examined whether the respiratory defect phenotypes of rsc mutants could be relieved by the overexpression of HAP4. For this experiment, we constructed a high-copy plasmid carrying HAP4 that was expressed under the control of the GPD promoter (pRS426GPDpr::HAP4). As shown in Fig 5A and 5B, defective growth of nps1-13 and rsc2Δ on YPEG plates and enhanced formation of petite nps1-13 colonies were alleviated by the overexpression of HAP4. In contrast, little recovery was observed with regard to the increased accumulation of ROS in nps1-13 cells (Fig 1E). These results suggest the involvement of RSC in the transcriptional activation of a set of respiratory genes, together with the HAP complex. However, the results also indicate that RSC might interact with factors other than the HAP complex to regulate mitochondrial function.


RSC Chromatin-Remodeling Complex Is Important for Mitochondrial Function in Saccharomyces cerevisiae.

Imamura Y, Yu F, Nakamura M, Chihara Y, Okane K, Sato M, Kanai M, Hamada R, Ueno M, Yukawa M, Tsuchiya E - PLoS ONE (2015)

Overexpression of HAP4 alleviated the respiratory defect of nps1-13.(A) Effect of HAP4 overexpression on nps1-13 growth on YPEG. WT (BY4743) and nps1-13 (BYI-3) cells harboring pRS426 (WT/v and nps1-13/v, respectively) or pRS426GPDpr::HAP4 (WT/HAP4 and nps1-13/HAP4, respectively) were grown to log phase in SD-Ura medium, spotted on YPD and YPEG plates in serial five-fold dilutions and incubated at 30°C for 3 days. (B) Effect of HAP4 overexpression on petit nps1-13 colony formation. The strains described in (A) were plated on YPEG; three independent colonies were subsequently picked, pre-cultured in SD-Ura medium, and separately grown in YPD medium. On the indicated days, 200 cells from each culture were plated on YPD plates and incubated at 30°C for 3 days. Data are presented as the means ± SEM (n = 3).
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Related In: Results  -  Collection

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pone.0130397.g005: Overexpression of HAP4 alleviated the respiratory defect of nps1-13.(A) Effect of HAP4 overexpression on nps1-13 growth on YPEG. WT (BY4743) and nps1-13 (BYI-3) cells harboring pRS426 (WT/v and nps1-13/v, respectively) or pRS426GPDpr::HAP4 (WT/HAP4 and nps1-13/HAP4, respectively) were grown to log phase in SD-Ura medium, spotted on YPD and YPEG plates in serial five-fold dilutions and incubated at 30°C for 3 days. (B) Effect of HAP4 overexpression on petit nps1-13 colony formation. The strains described in (A) were plated on YPEG; three independent colonies were subsequently picked, pre-cultured in SD-Ura medium, and separately grown in YPD medium. On the indicated days, 200 cells from each culture were plated on YPD plates and incubated at 30°C for 3 days. Data are presented as the means ± SEM (n = 3).
Mentions: To understand the relationship between the RSC and HAP complex, we first determined whether the hap4Δ mutant exhibits similar phenotypes as those induced by mitochondrial dysfunction in rsc mutants. The hap4Δ mutant also exhibited growth defects on a medium containing a non-fermentable carbon source and a high frequency of mtDNA loss (S1 Fig). Next, we examined whether the respiratory defect phenotypes of rsc mutants could be relieved by the overexpression of HAP4. For this experiment, we constructed a high-copy plasmid carrying HAP4 that was expressed under the control of the GPD promoter (pRS426GPDpr::HAP4). As shown in Fig 5A and 5B, defective growth of nps1-13 and rsc2Δ on YPEG plates and enhanced formation of petite nps1-13 colonies were alleviated by the overexpression of HAP4. In contrast, little recovery was observed with regard to the increased accumulation of ROS in nps1-13 cells (Fig 1E). These results suggest the involvement of RSC in the transcriptional activation of a set of respiratory genes, together with the HAP complex. However, the results also indicate that RSC might interact with factors other than the HAP complex to regulate mitochondrial function.

Bottom Line: RSC (Remodel the Structure of Chromatin) is an ATP-dependent chromatin remodeling complex essential for the growth of Saccharomyces cerevisiae.The rsc2Δ phenotypes were relieved by RSC1 overexpression, indicating that the isoforms play a redundant role in respiratory growth.Genome-wide expression analysis in nps1-13 under respiratory conditions suggested that RSC regulates the transcription of some target genes of the HAP complex, a transcriptional activator of respiratory gene expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima, 739-8530, Japan.

ABSTRACT
RSC (Remodel the Structure of Chromatin) is an ATP-dependent chromatin remodeling complex essential for the growth of Saccharomyces cerevisiae. RSC exists as two distinct isoforms that share core subunits including the ATPase subunit Nps1/Sth1 but contain either Rsc1or Rsc2. Using the synthetic genetic array (SGA) of the non-essential mutation method, we screened for mutations exhibiting synthetic growth defects in combination with the temperature-sensitive mutant, nps1-105, and found connections between mitochondrial function and RSC. rsc mutants, including rsc1Δ, rsc2Δ, and nps1-13, another temperature-sensitive nps1 mutant, exhibited defective respiratory growth; in addition, rsc2Δ and nps1-13 contained aggregated mitochondria. The rsc2Δ phenotypes were relieved by RSC1 overexpression, indicating that the isoforms play a redundant role in respiratory growth. Genome-wide expression analysis in nps1-13 under respiratory conditions suggested that RSC regulates the transcription of some target genes of the HAP complex, a transcriptional activator of respiratory gene expression. Nps1 physically interacted with Hap4, the transcriptional activator moiety of the HAP complex, and overexpression of HAP4 alleviated respiratory defects in nps1-13, suggesting that RSC plays pivotal roles in mitochondrial gene expression and shares a set of target genes with the HAP complex.

No MeSH data available.


Related in: MedlinePlus