Snapshots of RNA polymerase III in action – A mini review

Wang, Qianmin; Daiß, Julia L; Xu, Youwei; Engel, Christoph

doi:10.1016/j.gene.2022.146282

Cited by 10 publications

(5 citation statements)

References 104 publications

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“…This may indicate a mechanism to prevent binding of unspecific DNA. Similar strategies are employed in RNAP I and III using intrinsic structural elements (specifically, the C-terminus of subunit RPC7 in RNAP III 75 or the DNA-loop within the largest subunit of RNAP I 6,7 ). RNAPs decorated with Spt4/5 might represent an RNAP mimicking ‘expander’ reservoir available in the cell that can quickly be activated.…”

Section: Discussionmentioning

confidence: 99%

Structural basis of archaeal RNA polymerase transcription elongation and Spt4/5 recruitment

Tarau,

Grünberger,

Pilsl

et al. 2024

Preprint

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Archaeal transcription is carried out by a multi-subunit RNA polymerase (RNAP) that is highly homologous in structure and function to eukaryotic RNAP II. Among the set of basal transcription factors, only Spt5 is found in all domains of life but Spt5 has been shaped during evolution, which is also reflected in the heterodimerization of Spt5 with Spt4 in Archaea and Eukaryotes. To unravel the mechanistic basis of Spt4/5 function in Archaea, we performed structure-function analyses using the archaeal transcriptional machinery ofPyrococcus furiosus(Pfu). We report single-particle cryo-electron microscopy reconstructions of apo RNAP and the archaeal elongation complex (EC) in the absence and presence of Spt4/5. Surprisingly,PfuSpt4/5 also binds the RNAP in the absence of nucleic acids in a distinct super-contracted conformation. We show that the RNAP clamp/stalk module exhibits conformational flexibility in the apo state of RNAP and that the enzyme contracts upon EC formation or Spt4/5 engagement. We furthermore identified a contact of the Spt5-NGN domain with the DNA duplex that stabilizes the upstream boundary of the transcription bubble and impacts Spt4/5 activityin vitro. This study, therefore, provides the structural basis for Spt4/5 function in archaeal transcription and reveals a potential role beyond the well-described support of elongation.

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Section: Discussionmentioning

confidence: 99%

Structural basis of archaeal RNA polymerase transcription elongation and Spt4/5 recruitment

Tarau,

Grünberger,

Pilsl

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…This may indicate a mechanism to prevent the binding of unspecific DNA. Similar strategies are employed in RNAP I and III using intrinsic structural elements (specifically, the C-terminus of subunit RPC7 in RNAP III ( 97 ) or the DNA-loop within the largest subunit of RNAP I ( 6 , 7 )). RNAPs decorated with Spt4/5 might represent a dormant RNAP reservoir available in the cell that can quickly be activated.…”

Section: Discussionmentioning

confidence: 99%

Structural basis of archaeal RNA polymerase transcription elongation and Spt4/5 recruitment

Tarău,

Grünberger,

Pilsl

et al. 2024

Nucleic Acids Research

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Archaeal transcription is carried out by a multi-subunit RNA polymerase (RNAP) that is highly homologous in structure and function to eukaryotic RNAP II. Among the set of basal transcription factors, only Spt5 is found in all domains of life, but Spt5 has been shaped during evolution, which is also reflected in the heterodimerization of Spt5 with Spt4 in Archaea and Eukaryotes. To unravel the mechanistic basis of Spt4/5 function in Archaea, we performed structure-function analyses using the archaeal transcriptional machinery of Pyrococcus furiosus (Pfu). We report single-particle cryo-electron microscopy reconstructions of apo RNAP and the archaeal elongation complex (EC) in the absence and presence of Spt4/5. Surprisingly, Pfu Spt4/5 also binds the RNAP in the absence of nucleic acids in a distinct super-contracted conformation. We show that the RNAP clamp/stalk module exhibits conformational flexibility in the apo state of RNAP and that the enzyme contracts upon EC formation or Spt4/5 engagement. We furthermore identified a contact of the Spt5-NGN domain with the DNA duplex that stabilizes the upstream boundary of the transcription bubble and impacts Spt4/5 activity in vitro. This study, therefore, provides the structural basis for Spt4/5 function in archaeal transcription and reveals a potential role beyond the well-described support of elongation.

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“…This process is crucial for precise gene transcription and protein synthesis [46,47]. Pol III, the largest RNA polymerase, is highly conserved across eukaryotic organisms [48]. Nguyen et al [49] discovered that weak or absent RPC4 expression caused the loss-of-function alleles in rice DGS1-nivara s and DGS2-T65 s , leading to hybrid incompatibility.…”

Section: Discussionmentioning

confidence: 99%

Screening of Sugarcane Proteins Associated with Defense against Leifsonia xyli subsp. xyli, Agent of Ratoon Stunting Disease

Zhang,

Liang,

Zhang

et al. 2024

Plants

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Sugarcane is the most important sugar crop and one of the leading energy-producing crops in the world. Ratoon stunting disease (RSD), caused by the bacterium Leifsonia xyli subsp. xyli, poses a huge threat to ratoon crops, causing a significant yield loss in sugarcane. Breeding resistant varieties is considered the most effective and fundamental approach to control RSD in sugarcane. The exploration of resistance genes forms the foundation for breeding resistant varieties through molecular technology. The pglA gene is a pathogenicity gene in L. xyli subsp. xyli, encoding an endopolygalacturonase. In this study, the pglA gene from L. xyli subsp. xyli and related microorganisms was analyzed. Then, a non-toxic, non-autoactivating pglA bait was successfully expressed in yeast cells. Simultaneously the yeast two-hybrid library was generated using RNA from the L. xyli subsp. xyli-infected sugarcane. Screening the library with the pglA bait uncovered proteins that interacted with pglA, primarily associated with ABA pathways and the plant immune system, suggesting that sugarcane employs these pathways to respond to L. xyli subsp. xyli, triggering pathogenicity or resistance. The expression of genes encoding these proteins was also investigated in L. xyli subsp. xyli-infected sugarcane, suggesting multiple layers of regulatory mechanisms in the interaction between sugarcane and L. xyli subsp. xyli. This work promotes the understanding of plant–pathogen interaction and provides target proteins/genes for molecular breeding to improve sugarcane resistance to L. xyli subsp. xyli.

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Snapshots of RNA polymerase III in action – A mini review

Cited by 10 publications

References 104 publications

Structural basis of archaeal RNA polymerase transcription elongation and Spt4/5 recruitment

Structural basis of archaeal RNA polymerase transcription elongation and Spt4/5 recruitment

Structural basis of archaeal RNA polymerase transcription elongation and Spt4/5 recruitment

Screening of Sugarcane Proteins Associated with Defense against Leifsonia xyli subsp. xyli, Agent of Ratoon Stunting Disease

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