Biochemical Characterization of Yeast Xrn1.

Abstract:

:Messenger RNA degradation is an important component of overall gene expression. During the final step of eukaryotic mRNA degradation, exoribonuclease 1 (Xrn1) carries out 5' → 3' processive, hydrolytic degradation of RNA molecules using divalent metal ion catalysis. To initiate studies of the 5' → 3' RNA decay machinery in our lab, we expressed a C-terminally truncated version of Saccharomyces cerevisiae Xrn1 and explored its enzymology using a second-generation, time-resolved fluorescence RNA degradation assay. Using this system, we quantitatively explored Xrn1's preference for 5'-monophosphorylated RNA substrates, its pH dependence, and the importance of active site mutations in the molecule's conserved catalytic core. Furthermore, we explore Xrn1's preference for RNAs containing a 5' single-stranded region both in an intermolecular hairpin structure and in an RNA-DNA hybrid duplex system. These results both expand and solidify our understanding of Xrn1, a centrally important enzyme whose biochemical properties have implications in numerous RNA degradation and processing pathways.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Langeberg CJ,Welch WRW,McGuire JV,Ashby A,Jackson AD,Chapman EG

doi

10.1021/acs.biochem.9b01035

subject

Has Abstract

pub_date

2020-04-21 00:00:00

pages

1493-1507

issue

15

eissn

0006-2960

issn

1520-4995

journal_volume

59

pub_type

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