Open AccessTranscription-Dependent Formation of Nuclear Granules Containing FUS and RNA Pol II
Author(s) -
Valery F. Thompson,
Rachel A. Victor,
Andrés A. Morera,
Mahta Moinpour,
Meilani N. Liu,
Conner C. Kisiel,
Kaitlyn Pickrel,
Charis E. Springhower,
Jacob C. Schwartz
Publication year - 2018
Publication title -
biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.43
H-Index - 253
eISSN - 1520-4995
pISSN - 0006-2960
DOI - 10.1021/acs.biochem.8b01097
Subject(s) - rna polymerase ii , transcription (linguistics) , rna , microbiology and biotechnology , polymerase , chemistry , rna polymerase , rna polymerase i , processivity , rna dependent rna polymerase , transcription factor ii d , rna polymerase iii , in vitro , biology , biochemistry , gene expression , enzyme , gene , promoter , linguistics , philosophy
Purified recombinant FUsed in Sarcoma (FUS) assembles into an oligomeric state in an RNA-dependent manner to form large condensates. FUS condensates bind and concentrate the C-terminal domain of RNA polymerase II (RNA Pol II). We asked whether a granule in cells contained FUS and RNA Pol II as suggested by the binding of FUS condensates to the polymerase. We developed cross-linking protocols to recover protein particles containing FUS from cells and separated them by size exclusion chromatography. We found a significant fraction of RNA Pol II in large granules containing FUS with diameters of >50 nm or twice that of the RNA Pol II holoenzyme. Inhibition of transcription prevented the polymerase from associating with the granules. Altogether, we found physical evidence of granules containing FUS and RNA Pol II in cells that possess properties comparable to those of in vitro FUS condensates.