Mutational analysis of the RNA helicase Dhh1 in Ste12 expression and yeast mating

Daehee Jung; Jihye Ahn; Boram Rhee; Jinmi Kim

doi:10.1007/s12275-017-7020-4

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Mutational analysis of the RNA helicase Dhh1 in Ste12 expression and yeast mating: Daehee Jung , Jihye Ahn , Boram Rhee , Jinmi Kim; Journal of Microbiology 2017;55(5):373-378.
DOI: https://doi.org/10.1007/s12275-017-7020-4
Published online: April 29, 2017

Department of Microbiology and Molecular Biology, College of Bioscience and Biotechnology, Chungnam National University, Daejeon 34134, Republic of Korea

Corresponding author: Jinmi Kim , Tel: +82-42-821-6416,

Received: 11 January 2017 • Revised: 6 April 2017 • Accepted: 11 April 2017

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Abstract

Dhh1 and Dhh1 homologues (RCK/p54/DDX6) are mem-bers of the DEAD-box protein family of RNA helicases. These proteins display conserved sequence motifs for ATPase and RNA binding activities. Dhh1 is a component of the P-bodies (processing bodies) of mRNA granules and functions as an mRNA decapping activator in Saccharomyces cerevisiae. Dhh1 also contributes to gene-specific regulation during yeast mating. The dhh1 deletion mutation results in a significant decrease in the expression of Ste12, a mating-specific trans-cription factor, showing severe mating defects. Here, we in-troduced amino-acid substitution mutations in the ATPase and RNA binding domains of Dhh1 and also constructed a deletion of 79 amino acids at the Q/P-rich C-terminal region. The mutations in ATPase A and B motif (K96R, D195A) and C-terminus deletion showed reduced levels of mating effi-ciency as well as Ste12 protein expression. The Q/P-rich C- terminal region of Dhh1 was dispensable for growth at non- permissive temperature 37°C but appeared to play an im-portant role in regulating the Ste12 protein expression and mating processes. The P-body accumulation induced by treatment with α-mating factor required ATPase, RNA-bind-ing and the Q/P-rich C-terminal domains of Dhh1.

Keywords: RNA helicase;Dhh1;Ste12 expression;P-bodies;ATPase domain;Q/P-rich region

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Mutational analysis of the RNA helicase Dhh1 in Ste12 expression and yeast mating

J. Microbiol. 2017;55(5):373-378. Published online April 29, 2017

DOI: https://doi.org/10.1007/s12275-017-7020-4

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