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Investigating the role of Hmo1 in transcription by Pol I in yeast.

ABSTRACT: Investigating the role of Hmo1 in transcription by Pol I in yeast.

PROVIDER: PRJNA1041359 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR26849336.fastq.gz	Fastqsanger.gz
	SRR26849337.fastq.gz	Fastqsanger.gz
	SRR26849338.fastq.gz	Fastqsanger.gz

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Investigating the role of Hmo1 in transcription by Pol I in yeast.

Project description:The purpose of this study was to investigate the role of the yeast protein, high mobility group protein (Hmo1), in rRNA synthesis. For over twenty years, Hmo1 has been proposed to be an activator of transcription by Pol I, but its regulatory mechanism remains largely undefined. Therefore, we used native elongating transcript sequencing (NET-seq) to explore the role of this factor in transcription by Pol I. Our NET-seq results demonstrated that in cells lacking Hmo1 (hmo1D), Pol I occupancy was significantly increased across the rDNA template, indicating an increased pause propensity in these cells. These data suggest that Hmo1 is an important transcription elongation factor for Pol I.

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Project description:Polymerase-associated factor 1 complex (Paf1C) has been identified to promote transcription by Pols I and II and is conserved across eukaryotes. This transcription factor has been well-studied for Pol II, but its role in transcription by Pol I has not been fully characterized. Therefore, the goal of this study was to use a high-resolution technique, native elongating transcript sequencing (NET-seq), to investigate the effect of the loss of two of the five Paf1C subunits (Paf1 and Cdc73) on Pol I occupancy in yeast. Most notably, we found that in both paf1D and cdc73D mutants, there was a significant reduction in Pol I occupancy at the 5’ end of the rDNA template as compared to WT yeast. Overall, our results suggest that Paf1C is an important transcription elongation factor for Pol I.

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Project description:DNA topoisomerase-2 and high mobility group protein Hmo1 are known to regulate chromatin architecture by regulating gene boundaries. Here we report how these proteins affect global RNA level after inactivation of Top2 and Hmo1. Our data indicate that inactivating Hmo1 has a drastic effect on transcription levels of 20% yeast genes, however, this phenomenon can slightly be rescued by inactivating Top2 functions. Also, we study the Top2 and Top1 role in nucleosome architecture with and without expressing E.coli TopA. In top2-1;top1∆ condition with TopA expressed, it affects the nucleosome occupancy at the global level compared with top2-1;top1∆-Control plasmid. The ChIA-PET (Chromatin interaction analysis by paired-end tag sequencing) method is used to address whether a specific protein is engaged in the chromosomal interactions. Epitope tagged Top2 protein is used as probe in ChIA-PET experiments to map Top2 mediated chromatin-chromatin interactions.

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