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Investigating Conditional Mutations in MTR4 : an RNA Helicase Associated with the Nuclear Exosome
Author(s) -
Isenhour Anthony,
Fulop Hannah,
Pingali Hema,
Hilliker Angie
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.459.4
Subject(s) - biology , rna , mutagenesis , mutant , small nuclear rna , exosome complex , rna helicase a , genetics , microbiology and biotechnology , rna splicing , ribosomal rna , gene , helicase , non coding rna
The nuclear exosome is a complex of proteins in the nucleus that processes and degrades various RNAs. In doing so, it associates with various other proteins, including Mtr4, an RNA helicase that processes and degrades RNA. MTR4 is an essential gene conserved from yeast to humans. Mtr4 associates with various complexes, such as TRAMP and the nuclear exosome, to degrade aberrant RNAs and process RNAs such as the 5.8s rRNA and the U4 snRNA (1, 2). The ATPase region of Mtr4 has been shown to be essential, but the N‐ and C‐termini are not fully characterized (3). Current questions about MTR4 focus on how Mtr4 chooses which RNAs to process and degrade, as well as what regions of the protein affect said functions. Through random mutagenesis of the N‐ and C‐ termini of Mtr4, we selected for conditional mutants that are phenotypically defective only at high temperature. Because Mtr4 is essential, conditional mutants serve as in vivo tools to investigate how minor alterations to MTR4's protein sequence affect the cell. We have generated five C‐terminal conditional mutants, three N‐terminal conditional mutants, and a large library of lethal mutants. Through chemiluminescent northern blotting we have been able to confirm that our C‐terminal mutants are not defective in 5.8s rRNA processing. N‐terminal mutants testing is still underway, as well as U4 processing tests. Understanding the integral residues of Mtr4 will allow us to greater understand how it processes some RNAs and degrades others. Support or Funding Information NIH R15 GM128068‐01; Arts & Sciences (UR) This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .