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Genome-wide maps of RNA Pol III subunits POLR3C, POLR3G, and PLOR3GL in ES-E14TG2a and primary MEF cells

ABSTRACT: Genome-wide maps of RNA Pol III subunits POLR3C, POLR3G, and PLOR3GL in ES-E14TG2a and primary MEF cells

PROVIDER: PRJNA602395 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10918010.fastq.gz	Fastqsanger.gz
	SRR10918011.fastq.gz	Fastqsanger.gz
	SRR10918012.fastq.gz	Fastqsanger.gz
	SRR10918013.fastq.gz	Fastqsanger.gz
	SRR10918014.fastq.gz	Fastqsanger.gz

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Genome-wide maps of RNA Pol III subunits POLR3C, POLR3G, and PLOR3GL in ES-E14TG2a and primary MEF cells

Project description:Mammalian cells contain two isoforms of RNA Polymerase III (Pol III) that differ in only a single subunit, with POLR3G in one form and the related POLR3GL in the other form. Previous research indicates that POLR3G and POLR3GL are differentially expressed, with POLR3G expression being largely restricted to embryonic stem cells (ESCs) and tumor cells in contrast to the ubiquitously expressed POLR3GL. To date, the functional differences between these two subunits remain largely unexplored, especially in vivo. Here, we show that POLR3G and POLR3GL containing Pol III complexes bind the same target genes and assume the same functions both in vitro and in vivo and, to a certain degree, can compensate for each other in vivo. Moreover, an observed defect in the differentiation ability of POLR3G knockout ESCs can be rescued by exogenous expression of POLR3GL. Interestingly, POLR3G knockout mice die at a very early embryonic stage, whereas POLR3GL knockout mice complete embryonic development without noticeable defects but die at about 3 weeks after birth with signs of both general growth defects and potential cerebellum-related neuronal defects. The different phenotypes of the knockout mice likely reflect differential expression levels of POLR3G and POLR3GL across developmental stages and between tissues and insufficient amounts of total Pol III in vivo.

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Project description:Pol II has been recognized as a passively regulated holoenzyme. However, whether Pol II plays specific regulatory roles remain unclear. Here, fractions containing disassociated RPB3 (dRPB3) were identified by size exclusion chromatography in various cells. Through a unique strategy, Specific Degradation of Disassociated Subunits (SDDS), we demonstrated that dRPB3 functions as a regulatory component of Pol II to enable the preferential control of 3’ end processing of ribosomal protein genes directly through its N-terminal domain. Machine learning analysis of large-scale genomic features revealed that the little elongation complex helps to specialize the functions of dRPB3. Mechanistically, dRPB3 facilitates CBC-PCF11 axis activity to increase the efficiency of 3’ end processing. Furthermore, RPB3 is dynamically regulated during development and diseases. These findings suggest that Pol II gains specific regulatory functions by trapping disassociated subunits.

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