Assembly and isolation of intermediate steps of transcription complexes formed on the human 5S rRNA gene

Weser, Stephan; Riemann, Jana; Seifart, Klaus H.; Meißner, Wolfgang

doi:10.1093/nar/gkg345

Cited by 6 publications

(5 citation statements)

References 33 publications

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“…UCSC ribosome profiling data supports (almost) sole CUG initiation of GTF3A, R3HCC1 and BAG1 (Figure 5A – C top). GTF3A encodes the general transcription factor 3A, responsible for RNA polymerase III transcription ( 42 ). Intriguingly, its translation starts from a CUG codon in mammals, but in birds, frogs, fish, insects or lower eukaryotes, initiation occurs from the AUG codon located at the position equivalent to the CUG codon ( Supplementary Figure S6 ).…”

Section: Resultsmentioning

confidence: 99%

Competition between translation initiation factor eIF5 and its mimic protein 5MP determines non-AUG initiation rate genome-wide

Tang

Morris

Wan

et al. 2017

Nucleic Acids Research

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In the human genome, translation initiation from non-AUG codons plays an important role in various gene regulation programs. However, mechanisms regulating the non-AUG initiation rate remain poorly understood. Here, we show that the non-AUG initiation rate is nearly consistent under a fixed nucleotide context in various human and insect cells. Yet, it ranges from <1% to nearly 100% compared to AUG translation, depending on surrounding sequences, including Kozak, and possibly additional nucleotide contexts. Mechanistically, this range of non-AUG initiation is controlled in part, by the eIF5-mimic protein (5MP). 5MP represses non-AUG translation by competing with eIF5 for the Met-tRNAi-binding factor eIF2. Consistently, eIF5 increases, whereas 5MP decreases translation of NAT1/EIF4G2/DAP5, whose sole start codon is GUG. By modulating eIF5 and 5MP1 expression in combination with ribosome profiling we identified a handful of previously unknown non-AUG initiation sites, some of which serve as the exclusive start codons. If the initiation rate for these codons is low, then an AUG-initiated downstream ORF prevents the generation of shorter, AUG-initiated isoforms. We propose that the homeostasis of the non-AUG translatome is maintained through balanced expression of eIF5 and 5MP.

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Section: Resultsmentioning

confidence: 99%

Competition between translation initiation factor eIF5 and its mimic protein 5MP determines non-AUG initiation rate genome-wide

Tang

Morris

Wan

et al. 2017

Nucleic Acids Research

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“…TFIIIB␤ was further purified by chromatography over MonoQ (11), which also separated TFIIIB␤ (elution from 280 to 300 mM KCl) from TFIIIC1Ј (elution from 200 to 250 mM KCl). TFIIIB␤ activity co-eluted with HsTBP and HsBrf1 on MonoQ but was separated from any protein showing cross-reactivity with anti-HsBdp1 antibodies (data not shown).…”

Section: Methodsmentioning

confidence: 99%

“…It is essential not only for the transcription of type 1 and type 2 genes but also for that of type 3 genes and, thus, for all known human genes transcribed by RNA polymerase III (6,8,9). TFIIIC1 is not an assembly factor for TFIIIB␤ (defined below) but is required for the recruitment and initiation of RNA polymerase III (10,11). Also identified are two other activities, TFIIIC1-like and TFIIIC1Ј, that are functionally similar to TFIIIC1 but are chromatographically distinct (9,(12)(13)(14).…”

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confidence: 99%

Transcription Factor (TF)-like Nuclear Regulator, the 250-kDa Form of Homo sapiens TFIIIB″, Is an Essential Component of Human TFIIIC1 Activity

Weser

Gruber

Häfner

et al. 2004

Journal of Biological Chemistry

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The general human RNA polymerase III transcription factor (TF) IIIC1 has hitherto been ill defined with respect to the polypeptides required for reconstitution of its activity. Here we identify Homo sapiens TFIIIB؆ (HsBdp1) as an essential component of hTFIIIC1 and hT-FIIIC1-like activities. Several forms of HsBdp1 are described. The 250-kDa form of HsBdp1, also designated the "transcription factor-like nuclear regulator," strictly co-eluted with TFIIIC1 activity over multiple chromatographic purification steps as revealed by Western blot with anti-HsBdp1 antibodies and by MALDI-TOF analysis. In addition, TFIIIC1 activity could be depleted from partially purified fractions with antiHsBdp1 antibodies but not with control antibodies. Moreover, highly purified recombinant HsBdp1 could replace TFIIIC1 activity in reconstituted transcription of the VAI gene in vitro. Furthermore, smaller proteins of ϳ90 -150 kDa that were recognized by anti-HsBdp1 antibodies co-eluted with TFIIIC1-like activity. Finally, cytoplasmic extracts from differentiated mouse F9 fibroblast cells that lacked TFIIIC1 activity could be made competent for transcription of the VA1 gene by the addition of TFIIIC1, TFIIIC1-like, or recombinant HsBdp1. These results suggest that HsBdp1 proteins represent essential components of TFIIIC1 and TFIIIC1-like activities.RNA polymerase III transcribes genes encoding small, untranslated RNAs including tRNA, 5 S rRNA, and U6 small nuclear RNA genes (reviewed in Ref. 1). In the yeast Saccharomyces cerevisiae, genes transcribed by RNA polymerase III are governed by promoter elements comprised of A and C boxes (type 1) or A and B boxes (type 2) that are located downstream of the transcription initiation site. Two transcription factors (TF), 1 TFIIIB and TFIIIC, are necessary and sufficient for the transcription of type 2 genes (tRNA), whereas transcription of the type 1 5 S rRNA gene requires, in addition, TFIIIA. S. cerevisiae (Sc) TFIIIC is a stable complex of six polypeptides, whereas ScTFIIIB consists of a less stable association of three components, namely the ScTBP, ScBrf1, and ScBdp1 proteins (reviewed in Refs.

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“…Transcriptional factor III A (GTF3A), an RNA polymerase III transcriptional factor, specifically binds to the internal control region of the 5S rRNA gene from nucleotides +43 to +96[ 3 ], initiating the assembly of the transcription initiation complex (5S rDNA-TFIIIA-TFIIIC2-TFIIIBbeta complex). Complex formation is not proportional to the amount of TFIIIA[ 4 ]. GTF3A is present in all human organs.…”

Section: Introductionmentioning

confidence: 99%

Transcriptional factor III A promotes colorectal cancer progression by upregulating cystatin A

Wang¹,

Yuan²,

Jia³

et al. 2022

World J Gastrointest Oncol

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BACKGROUND Advanced colorectal cancer (CRC) generally has poor outcomes and high mortality rates. Clarifying the molecular mechanisms underlying CRC progression is necessary to develop new diagnostic and therapeutic strategies to improve CRC outcome and decrease mortality. Transcriptional factor III A (GTF3A), an RNA polymerase III transcriptional factor, is a critical driver of tumorgenesis and aggravates CRC cell growth. AIM To confirm whether GTF3A promotes CRC progression by regulating the expression of cystatin A ( Csta ) gene and investigate whether GTF3A can serve as a prognostic biomarker and therapeutic target for patients with CRC. METHODS Human tissue microarrays containing 90 pairs of CRC tissues and adjacent non-tumor tissues, and human tissue microarrays containing 20 pairs of CRC tissues, adjacent non-tumor tissues, and metastatic tissues were examined for GTF3A expression using immunohistochemistry. The survival rates of patients were analyzed. Short hairpin GTF3As and CSTAs were designed and packaged into the virus to block the expression of Gtf3a and Csta genes, respectively. In vivo tumor growth assays were performed to confirm whether GTF3A promotes CRC cell proliferation in vivo. Electrophoretic mobility shift assay and fluorescence in situ hybridization assay were used to detect the interaction of GTF3A with Csta, whereas luciferase activity assay was used to evaluate the expression of the Gtf3a and Csta genes. RNA-Sequencing (RNA-Seq) and data analyses were used to screen for target genes of GTF3A. RESULTS The expression of GTF3A was higher in CRC tissues and lymph node metastatic tissues than in the adjacent normal tissues. GTF3A was associated with CRC prognosis, and knockdown of the Gtf3a gene impaired CRC cell proliferation, invasion, and motility in vitro and in vivo . Moreover, RNA-Seq analysis revealed that GTF3A might upregulate the expression of Csta , whereas the luciferase activity assay showed that GTF3A bound to the promoter of Csta gene and increased Csta transcription. Furthermore, CSTA regulated the expression of epithelial-mesenchymal transition (EMT) markers. CONCLUSION GTF3A increases CSTA expression by binding to the Csta promoter, and increased CSTA level promotes CRC progression by regulating the EMT. Inhibition of GTF3A prevents CRC progression. Therefore, GTF3A is a potential novel therapeutic target and biomarker for CRC.

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Assembly and isolation of intermediate steps of transcription complexes formed on the human 5S rRNA gene

Cited by 6 publications

References 33 publications

Competition between translation initiation factor eIF5 and its mimic protein 5MP determines non-AUG initiation rate genome-wide

Competition between translation initiation factor eIF5 and its mimic protein 5MP determines non-AUG initiation rate genome-wide

Transcription Factor (TF)-like Nuclear Regulator, the 250-kDa Form of Homo sapiens TFIIIB″, Is an Essential Component of Human TFIIIC1 Activity

Transcriptional factor III A promotes colorectal cancer progression by upregulating cystatin A

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