Fructose bisphosphate aldolase is involved in the control of RNA polymerase III-directed transcription

Cieśla, Małgorzata; Mierzejewska, Jolanta; Adamczyk, M.; Farrants, Ann‐Kristin Östlund; Boguta, Magdalena

doi:10.1016/j.bbamcr.2014.02.007

Cited by 38 publications

(29 citation statements)

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“…The growth of transformants was tested on glucose or glycerol medium at temperatures suitable for the individual mutants (35). b The complementation of Pol III mutants by increasing the gene dosage is indicated by ϩ or Ϫ. NT, not tested.…”

Section: Resultsmentioning

confidence: 99%

“…RNA was hybridized as described previously (35). The probes were 5=-GCGCTCTCCCAACTGAGCT-3= for tRNAPhe (GAA), 5=-TATTCCCACAGTTAACTGCGGTCA-3= for the intron of tRNA-Leu (CCA), 5=-CGAGTCGAACGCCCGAT-3= for tRNA-Tyr (GUA), and 5=-GCGTTGTTCATCGATGC-3= for 5.8S rRNA.…”

Section: Strainsmentioning

confidence: 99%

“…Immunofluorescence was performed as described previously (35). 9E10 anti-Myc mouse antibody (1:500 for 1 h; Roche) and anti-HA mouse antibody (1:500 for 1 h; Covance) were added, followed by secondary Cy3-conjugated anti-mouse antibody (1: 250 for 1 h; Jackson ImmunoResearch Laboratories).…”

Section: Western Analysismentioning

confidence: 99%

“…Western blotting was performed as described previously (35). Protein extracts were separated by 8%, 10%, or 12% SDS-PAGE and hybridized with mouse monoclonal antibodies anti-HA (Covance) at a 1:5,000 dilution for 2 h, anti-GFP (Roche) at a 1:5,000 dilution for 1 h, 9E10 anti-Myc (Roche) at a 1:2,000 dilution for 1 h, and antiVma2 (Molecular Probes) at a 1:10,000 dilution for 1 h and with rabbit polyclonal antibodies anti-HA (Sigma) at a 1:2,000 dilution for 1 h, antiRpb1 (Santa Cruz Biotechnology) at a 1:1,000 dilution for 1 h, anti-C82 at a 1:5,000 dilution for 1 h, anti-C53 at a 1:20,000 dilution for 1 h, and anti-AC40 at a 1:1,000 dilution for 2 h. Antibodies specific for Pol III subunits were a gift from O. Lefebvre.…”

Section: Western Analysismentioning

confidence: 99%

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Rbs1, a New Protein Implicated in RNA Polymerase III Biogenesis in Yeast Saccharomyces cerevisiae

Cieśla

Makała

Płonka

et al. 2015

Molecular and Cellular Biology

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Little is known about the RNA polymerase III (Pol III) complex assembly and its transport to the nucleus. We demonstrate that a missense cold-sensitive mutation, rpc128-1007, in the sequence encoding the C-terminal part of the second largest Pol III subunit, C128, affects the assembly and stability of the enzyme. The cellular levels and nuclear concentration of selected Pol III subunits were decreased in rpc128-1007 cells, and the association between Pol III subunits as evaluated by coimmunoprecipitation was also reduced. To identify the proteins involved in Pol III assembly, we performed a genetic screen for suppressors of the rpc128-1007 mutation and selected the Rbs1 gene, whose overexpression enhanced de novo tRNA transcription in rpc128-1007 cells, which correlated with increased stability, nuclear concentration, and interaction of Pol III subunits. The rpc128-1007 rbs1⌬ double mutant shows a synthetic growth defect, indicating that rpc128-1007 and rbs1⌬ function in parallel ways to negatively regulate Pol III assembly. Rbs1 physically interacts with a subset of Pol III subunits, AC19, AC40, and ABC27/Rpb5. Additionally, Rbs1 interacts with the Crm1 exportin and shuttles between the cytoplasm and nucleus. We postulate that Rbs1 binds to the Pol III complex or subcomplex and facilitates its translocation to the nucleus.A ll eukaryotic cells have at least three different RNA polymerases (Pol). Pol I synthesizes the large precursor of rRNA, Pol II produces mainly mRNAs, and Pol III generates tRNAs, 5S rRNA and other small noncoding RNAs.Pol III, which is the focus of this work, is a heteromultimeric protein complex that contains 17 distinct subunits in Saccharomyces cerevisiae. The two largest subunits, C160 and C128, form opposite sides of the active-center cleft and harbor the catalytic activity, and they are related to the ␤= and ␤ components of the ␣ 2 ␤␤= core of bacterial RNA polymerase. Homology to the bacterial ␣ subunit, although less strong, was also observed, with the subunit AC40 common for yeast and Pol III and Pol I. A second ␣-like subunit, AC19, forms a heterodimer with AC40, which is a functional equivalent to the ␣ 2 homodimer in prokaryotes (1). The polymerase core, in addition to the ␣ 2 ␤␤=-like structure, contains six small subunits, which are structurally and functionally conserved from yeast to humans. The small core subunits either bind or bridge the two largest catalytic subunits. In addition to the central core, the remaining subunits of Pol III are organized in heterodimeric or trimeric stalks or subdomains. Significantly, the C37 to C53 and C82 to C34 subdomains, which are stably bound to the RNA Pol III core, resemble Pol II general transcription factors TFIIF and TFIIE (2).Little is known about how polymerase complexes are assembled from the subunits, how they reach their functional destination, and how they dissociate after transcription. In bacteria, the assembly of RNA polymerase starts with the formation of the ␣␣ dimer, which then interacts with the ␤ subunit to yield an ␣␣␤...

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Section: Resultsmentioning

confidence: 99%

Section: Strainsmentioning

confidence: 99%

Section: Western Analysismentioning

confidence: 99%

Section: Western Analysismentioning

confidence: 99%

See 2 more Smart Citations

Rbs1, a New Protein Implicated in RNA Polymerase III Biogenesis in Yeast Saccharomyces cerevisiae

Cieśla

Makała

Płonka

et al. 2015

Molecular and Cellular Biology

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“…All isoforms have been found in the nucleus, where they bind to DNA and interact with RNA polymerase III [31][32][33]. The nuclear translocation of ALDO is associated with increased proliferation but its precise role is unknown [34].…”

Section: Glycolytic Enzymes In the Nucleusmentioning

confidence: 98%

Metabolic Enzymes Moonlighting in the Nucleus: Metabolic Regulation of Gene Transcription

Boukouris

Zervopoulos

Michelakis

2016

Trends in Biochemical Sciences

240

206

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Intensive herbicide use has selected for constitutively elevated levels of stress‐responsive mRNAs and proteins in multiple herbicide‐resistant Avena fatua L

Keith

Burns

Bothner

et al. 2017

Pest Management Science

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Fructose bisphosphate aldolase is involved in the control of RNA polymerase III-directed transcription

Cited by 38 publications

References 50 publications

Rbs1, a New Protein Implicated in RNA Polymerase III Biogenesis in Yeast Saccharomyces cerevisiae

Rbs1, a New Protein Implicated in RNA Polymerase III Biogenesis in Yeast Saccharomyces cerevisiae

Metabolic Enzymes Moonlighting in the Nucleus: Metabolic Regulation of Gene Transcription

Intensive herbicide use has selected for constitutively elevated levels of stress‐responsive mRNAs and proteins in multiple herbicide‐resistant Avena fatua L

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