RNA polymerase II (RNAPII) is an essential machinery for catalyzing mRNA synthesis and controlling cell fate in eukaryotes. Although the structure and function of RNAPII have been relatively defined, the molecular mechanism of its assembly process is not clear. The identification and functional analysis of assembly factors will provide new understanding to transcription regulation. In this study, we identify that RTR1, a known transcription regulator, is a new multicopy genetic suppressor of mutants of assembly factors Gpn3, Gpn2, and Rba50. We demonstrate that Rtr1 is directly required to assemble the two largest subunits of RNAPII by coordinating with Gpn3 and Npa3. Deletion of RTR1 leads to cytoplasmic clumping of RNAPII subunit and multiple copies of RTR1 can inhibit the formation of cytoplasmic clump of RNAPII subunit in gpn3-9 mutant, indicating a new layer function of Rtr1 in checking proper assembly of RNAPII. In addition, we find that disrupted activity of Rtr1 phosphatase does not trigger the formation of cytoplasmic clump of RNAPII subunit in a catalytically inactive mutant of RTR1. Based on these results, we conclude that Rtr1 cooperates with Gpn3 and Npa3 to assemble RNAPII core.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1096/fj.202200698RR | DOI Listing |
J Microsc
December 2024
Cambridge Institute for Medical Research, University of Cambridge, Cambridge, UK.
Conventional optical microscopy imaging of obligate intracellular bacteria is hampered by the small size of bacterial cells, tight clustering exhibited by some bacterial species and challenges relating to labelling such as background from host cells, a lack of validated reagents, and a lack of tools for genetic manipulation. In this study, we imaged intracellular bacteria from the species Orientia tsutsugamushi (Ot) using five different fluorescence microscopy techniques: standard confocal, Airyscan confocal, instant Structured Illumination Microscopy (iSIM), three-dimensional Structured Illumination Microscopy (3D-SIM) and Stimulated Emission Depletion Microscopy (STED). We compared the ability of each to resolve bacterial cells in intracellular clumps in the lateral (xy) axis, using full width half-maximum (FWHM) measurements of a labelled outer membrane protein (ScaA) and the ability to detect small, outer membrane vesicles external to the cells.
View Article and Find Full Text PDFPlant Physiol
December 2024
Department of Plant and Environmental Sciences, Weizmann Institute of Science, Rehovot 76100, Israel.
Osmotic stress, caused by the lack of water or by high salinity, is a common problem in plant roots. Osmotic stress can be reproducibly simulated with the application of solutions of the high-molecular-weight and impermeable polyethylene glycol. The accumulation of different reactive oxygen species, such as singlet oxygen, superoxide, and hydrogen peroxide, accompany this stress.
View Article and Find Full Text PDFBackground Breast carcinoma is the most common cause of death in women arising primarily from either epithelial or stromal components. Primary breast lymphoma (PBL) is a less common type of neoplasm arising from the lymphoid tissue and is classified as non-Hodgkin's lymphoma (NHL). PBL is a rare tumor that accounts for less than 1% of total primary malignant neoplasms of the breast.
View Article and Find Full Text PDFMetabolism
January 2025
Kasturba Medical College, Manipal Academy of Higher Education, Manipal, Karnataka, India. Electronic address:
Conventional optical microscopy imaging of obligate intracellular bacteria is hampered by the small size of bacterial cells, tight clustering exhibited by some bacterial species and challenges relating to labelling such as background from host cells, a lack of validated reagents, and a lack of tools for genetic manipulation. In this study we imaged intracellular bacteria from the species (Ot) using five different fluorescence microscopy techniques: standard confocal, Airyscan confocal, instant Structured Illumination Microscopy (iSIM), three-dimensional Structured Illumination Microscopy (3D-SIM) and Stimulated Emission Depletion Microscopy (STED). We compared the ability of each to resolve bacterial cells in intracellular clumps in the lateral (xy) axis, using full width half maximum (FWHM) measurements of a labelled outer membrane protein (ScaA) and the ability to detect small, outer membrane vesicles external to the cells.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!