Zinc fingers 1 and 7 of yeast TFIIIA are essential for assembly of a functional transcription complex on the 5 S RNA gene

Rothfels, Karen; Rowland, Owen; Segall, J

doi:10.1093/nar/gkm517

Cited by 10 publications

(7 citation statements)

References 62 publications

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“…As a matter of fact, cells devoid of TFIIIA may survive if a single copy of the 5S rRNA gene is expressed from a RPR1 Pol III promoter, showing that the 5S rRNA transport and storage function of TFIIIA is dispensable in yeast. Consistent with this result is the survival of several mutants affecting the central fingers 4–6 shown to form the minimal 5S rRNA binding domain in Xenopus (27–29): disruption of yeast TFIIIA fingers 4, 5, 6, 4 + 5 or 4 + 6 yielded viable phenotypes (31). …”

Section: Introductionmentioning

confidence: 70%

Dicistronic tRNA–5S rRNA genes in Yarrowia lipolytica: an alternative TFIIIA-independent way for expression of 5S rRNA genes

Acker¹,

Ozanne²,

Kachouri‐Lafond³

et al. 2008

Nucleic Acids Research

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In eukaryotes, genes transcribed by RNA polymerase III (Pol III) carry their own internal promoters and as such, are transcribed as individual units. Indeed, a very few cases of dicistronic Pol III genes are yet known. In contrast to other hemiascomycetes, 5S rRNA genes of Yarrowia lipolytica are not embedded into the tandemly repeated rDNA units, but appear scattered throughout the genome. We report here an unprecedented genomic organization: 48 over the 108 copies of the 5S rRNA genes are located 3′ of tRNA genes. We show that these peculiar tRNA–5S rRNA dicistronic genes are expressed in vitro and in vivo as Pol III transcriptional fusions without the need of the 5S rRNA gene-specific factor TFIIIA, the deletion of which displays a viable phenotype. We also report the existence of a novel putative non-coding Pol III RNA of unknown function about 70 nucleotide-long (RUF70), the 13 genes of which are devoid of internal Pol III promoters and located 3′ of the 13 copies of the tDNA-Trp (CCA). All genes embedded in the various dicistronic genes, fused 5S rRNA genes, RUF70 genes and their leader tRNA genes appear to be efficiently transcribed and their products correctly processed in vivo.

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Section: Introductionmentioning

confidence: 70%

Dicistronic tRNA–5S rRNA genes in Yarrowia lipolytica: an alternative TFIIIA-independent way for expression of 5S rRNA genes

Acker¹,

Ozanne²,

Kachouri‐Lafond³

et al. 2008

Nucleic Acids Research

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“…The association of TF-IIIA represents the first step in the assembly of the pol III complex, by both inducing a minor bend in the DNA as well as assisting in the incorporation of TF-IIIC into the polymerase complex. TF-IIIB, in turn, induces a major bend in the DNA at the transcriptional start site [10,11]. Again, this complex formation is assisted by the association of MYC with TF-IIIB, in the absence of MAX, and the recruitment of the HAT complex, and either p53 or the pRB/p130 complex can suppress RNA pol III-mediated transcription (Figure 1) [8,9].…”

Section: Ribosome Biogenesis: An Overviewmentioning

confidence: 99%

Signal Transduction in Ribosome Biogenesis: A Recipe to Avoid Disaster

Piazzi

Bavelloni

Gallo

et al. 2019

IJMS

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Energetically speaking, ribosome biogenesis is by far the most costly process of the cell and, therefore, must be highly regulated in order to avoid unnecessary energy expenditure. Not only must ribosomal RNA (rRNA) synthesis, ribosomal protein (RP) transcription, translation, and nuclear import, as well as ribosome assembly, be tightly controlled, these events must be coordinated with other cellular events, such as cell division and differentiation. In addition, ribosome biogenesis must respond rapidly to environmental cues mediated by internal and cell surface receptors, or stress (oxidative stress, DNA damage, amino acid depletion, etc.). This review examines some of the well-studied pathways known to control ribosome biogenesis (PI3K-AKT-mTOR, RB-p53, MYC) and how they may interact with some of the less well studied pathways (eIF2α kinase and RNA editing/splicing) in higher eukaryotes to regulate ribosome biogenesis, assembly, and protein translation in a dynamic manner.

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“…Because PSTVd replication from circular genomic RNA to (-) oligomers and then to plus-strand or (+) oligomers is a continuous process, failure to tease apart each step resulted in controversial data, as evidenced by previous reports [34,41]. To understand whether Pol II can catalyze transcription using (-) viroid oligomers, we established a reconstituted in vitro transcription system using partially purified Pol II from wheat germ [24, 29] and the (-) PSTVd dimer as RNA template.…”

Section: Resultsmentioning

confidence: 99%

“…TFIIIA-7ZF has seven C2H2 type zinc finger domains. We mutated each zinc finger domain by changing the first histidine in the C2H2 domain to asparagine, which is commonly used to disrupt the local structure of a C2H2 motif [33, 34]. We then used those variants for the IVT assay.…”

Section: Resultsmentioning

confidence: 99%

A remodeled RNA polymerase II complex catalyzing viroid RNA-templated transcription

Mudiyanselage

Pechan

et al. 2022

Preprint

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Viroids, a group of plant pathogens, are mysterious subviral agents composed of single-stranded circular noncoding RNAs. Nuclear-replicating viroids exploit host RNA polymerase II (Pol II) activity for transcription from circular RNA genome to minus-strand intermediates, a classic example illustrating the intrinsic RNA-dependent RNA polymerase activity of Pol II. The mechanism for Pol II to accept single-stranded RNAs as templates for transcription remains poorly understood. Here, we reconstituted a robust in vitro transcription system and demonstrated that Pol II also accepts the minus-strand viroid RNA template to generate sense-strand RNAs. Based on this reconstituted system, we purified the Pol II complex on RNA templates for nano-liquid chromatography-tandem mass spectrometry analysis. We identified a remodeled Pol II missing Rpb5, Rpb6, and Rpb9, which contrasts to the canonical 12-subunit Pol II or the 10-subunit Pol II core on DNA templates. This remodeled Pol II is active in transcription with the aid of TFIIIA-7ZF. Interestingly, this remodeled Pol II appears not to require other canonical general transcription factors (such as TFIIA, TFIIB, TFIID, TFIIE, TFIIF, TFIIH, and TFIIS), indicating a different mechanism/machinery regulating RNA-templated transcription. Transcription elongation factors, such as FACT complex, PAF1 complex, and SPT6, were also absent in the transcription complex on RNA templates. We further analyzed the critical zinc finger domains in TFIIIA-7ZF for aiding Pol II activity on RNA template and revealed the first three zinc finger domains pivotal for template binding. Collectively, our data illustrated a distinct organization of Pol II complex on viroid RNA templates, providing new insights into the evolution of transcription machinery, the mechanism of RNA-templated transcription, as well as viroid replication.

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Zinc fingers 1 and 7 of yeast TFIIIA are essential for assembly of a functional transcription complex on the 5 S RNA gene

Cited by 10 publications

References 62 publications

Dicistronic tRNA–5S rRNA genes in Yarrowia lipolytica: an alternative TFIIIA-independent way for expression of 5S rRNA genes

Dicistronic tRNA–5S rRNA genes in Yarrowia lipolytica: an alternative TFIIIA-independent way for expression of 5S rRNA genes

Signal Transduction in Ribosome Biogenesis: A Recipe to Avoid Disaster

A remodeled RNA polymerase II complex catalyzing viroid RNA-templated transcription

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