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

• Published in: The journal of microbiology Vol. 55; no. 5; pp. 373 - 378
• Main Authors: Jung, Daehee, Ahn, Jihye, Rhee, Boram, Kim, Jinmi
• Format: Journal Article
• Language: English
• Published: Seoul The Microbiological Society of Korea 01.05.2017; Springer Nature B.V; 한국미생물학회
• Subjects: Adenosine Triphosphatases - genetics; Adenosine Triphosphatases - metabolism; adenosinetriphosphatase; Amino Acid Substitution; Amino acids; Biomedical and Life Sciences; DEAD-box RNA helicases; DEAD-box RNA Helicases - chemistry; DEAD-box RNA Helicases - genetics; DEAD-box RNA Helicases - metabolism; DNA Mutational Analysis; Gene Expression Regulation, Fungal; Genomics and Molecular Biology; granules; Life Sciences; messenger RNA; Microbial Genetics; Microbiology; Mutagenesis; Mutation; mutational analysis; Plasmids; Protein Biosynthesis; Protein expression; Protein Interaction Domains and Motifs; Protein Precursors - genetics; Protein Precursors - metabolism; protein synthesis; Proteins; RNA Recognition Motif Proteins - genetics; RNA Recognition Motif Proteins - metabolism; RNA, Messenger - genetics; Saccharomyces cerevisiae; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - physiology; Saccharomyces cerevisiae Proteins - chemistry; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; sequence deletion; temperature; Transcription factors; Transcription Factors - genetics; Transcription Factors - metabolism; Yeast; Yeasts; 생물학; P-bodies; Ste12 expression; ATPase domain; Q/P-rich region; Dhh1; RNA helicase
• ISSN: 1225-8873; 1976-3794; 1976-3794
• DOI: 10.1007/s12275-017-7020-4

Dhh1 and Dhh1 homologues (RCK/p54/DDX6) are members 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 transcription factor, showing severe mating defects. Here, we introduced 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 efficiency as well as Ste12 protein expression. The Q/P-rich C-terminal region of Dhh1 was dispensable for growth at nonpermissive temperature 37°C but appeared to play an important role in regulating the Ste12 protein expression and mating processes. The P-body accumulation induced by treatment with α-mating factor required ATPase, RNA-binding and the Q/P-rich C-terminal domains of Dhh1.