Journal of Microbiology

Journal Article

Mutational analysis of the RNA helicase Dhh1 in Ste12 expression and yeast mating: Daehee Jung , Jihye Ahn , Boram Rhee , Jinmi Kim; 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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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.

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Fus3 and Tpk2 protein kinases regulate the phosphorylation-dependent functions of RNA helicase Dhh1 in yeast mating and Ste12 protein expression
Jaehee Hwang, Daehee Jung, Jinmi Kim
Journal of Microbiology.2022; 60(8): 843. CrossRef
The Role of DEAD-Box ATPases in Gene Expression and the Regulation of RNA–Protein Condensates
Karsten Weis, Maria Hondele
Annual Review of Biochemistry.2022; 91(1): 197. CrossRef
Roles of Dhh1 RNA helicase in yeast filamentous growth: Analysis of N-terminal phosphorylation residues and ATPase domains
Eunji Lee, Daehee Jung, Jinmi Kim
Journal of Microbiology.2020; 58(10): 853. CrossRef
Functional association of Loc1 and Puf6 with RNA helicase Dhh1 in translational regulation of Saccharomyces cerevisiae Ste12
Daehee Jung, Jong Seok Seo, Jayoung Nam, Jinmi Kim, Enrico Baruffini
PLOS ONE.2019; 14(7): e0220137. CrossRef
Roles of eIF4E-binding protein Caf20 in Ste12 translation and P-body formation in yeast
Kiyoung Park, Yu-Seon Lee, Daehee Jung, Jinmi Kim
Journal of Microbiology.2018; 56(10): 744. CrossRef

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Identification of Psk2, Skp1, and Tub4 as trans-acting factors for uORF-containing ROK1 mRNA in Saccharomyces cerevisiae: Soonmee Jeon , Suran Lim , Jeemin Ha , Jinmi Kim; J. Microbiol. 2015;53(9):616-622. Published online August 27, 2015; DOI: https://doi.org/10.1007/s12275-015-5389-5

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Abstract

Rok1, a DEAD-box RNA helicase, is involved in rRNA processing and the control of cell cycle progression in Saccharomyces cerevisiae. Rok1 protein expression is cell cycle-regulated, declining at G1/S and increasing at G2. The downregulation of Rok1 expression in G1/S phase is mediated by the inhibitory action of two upstream open reading frames (uORFs) in the ROK1 5􍿁-untranslated region (5􍿁UTR). We identified Psk2 (PAS kinase), Skp1 (kinetochore protein) and Tub4 (γ-tubulin protein) as ROK1 5􍿁UTR-interacting proteins using yeast three-hybrid system. A deletion analysis of PSK2 or inactivation of temperature-sensitive alleles of SKP1 and TUB4 revealed that Rok1 protein synthesis is repressed by Psk2 and Skp1. This repression appeared to be mediated through the ROK1 uORF1. In contrast, Tub4 plays a positive role in regulating Rok1 protein synthesis and likely after the uORF1-mediated inhibitory regulation. These results suggest that 5􍿁UTR-interacting proteins, identified using three hybrid screening, are important for uORF-mediated regulation of Rok1 protein expression.

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Identification of short open reading frames in plant genomes
Yong Feng, Mengyun Jiang, Weichang Yu, Jiannan Zhou
Frontiers in Plant Science.2023;[Epub] CrossRef
HST1 increases replicative lifespan of a sir2Δ mutant in the absence of PDE2 in Saccharomyces cerevisiae
Woo Kyu Kang, Mayur Devare, Jeong-Yoon Kim
Journal of Microbiology.2017; 55(2): 123. CrossRef

Kaposi’s Sarcoma-Associated Herpesvirus Viral Protein Kinase Interacts with RNA Helicase A and Regulates Host Gene Expression: Jae Eun Jong , Junsoo Park , Sunmi Kim , Taegun Seo; J. Microbiol. 2010;48(2):206-212. Published online May 1, 2010; DOI: https://doi.org/10.1007/s12275-010-0021-1

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Abstract: RNA helicase A (RHA) containing the DExH motif is a human homolog of maleless protein that regulates expression of genes located in the Drosophila X chromosome during dosage compensation. RHA exerts helicase activity that unwinds double-stranded RNA and DNA to a single-strand form. The protein acts as a bridging factor mediating interactions of CBP/p300 and RNA pol II, and consequently affects gene expression. Kaposi’s sarcoma-associated herpesvirus (KSHV) is a member of the γ-herpesvirus subfamily that causes several disorders. The majority of herpesviruses commonly encode predicted viral protein kinases. KSHV open reading frame 36 (ORF36) codes for protein kinase domains, and functions as a serine/threonine protein kinase. KSHV ORF36 is classified as a late gene, as it is expressed during lytic replication and localized in the nuclei of KSHV-infected cells. Recent studies show that viral protein kinase (vPK) interacts with cellular proteins. In this study, we determined the cellular localization of vPK in KSHVinfected BCBL-1 cells using confocal microscopy. Proteomic analysis indicates that cellular proteins interacted with vPK, and co-immunoprecipitation reactions further reveal interactions between vPK and RHA. Moreover, KSHV vPK appeared to regulate the transcriptional activation of Cre promoter, and plays an important role in cellular transcription of RHA.

High Dosage of Rok1p, a Putative ATP-dependent RNA Helicase, Leads to a Cell Cycle Arrest at G1/S Stage in Saccharomyces cerevisiae: jeong, Hyun Sook , Oh, Jae Young , Kim, Jin Mi; J. Microbiol. 1998;36(2):139-144.

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Abstract: The ROK1 gene encodes a putative ATP-dependent RNA helicase which is essential for mitotic cell growth. ROK1 has been thought to affect microtubule and spindle pole body (SPB) functions in Saccharomyces cerevisiae. To investigate the intracellular functions of ROK1, we varied the Rok1 protein dosage in a cell and analyzed its phenotypic effects. Overexpression of the ROK1 gene by using a strong GAL1 promoter was lethal, leading cells to arrest at the unbudded stage. This arrest phenotype is very similar to that of the rok1 null mutation. Indirect immunofluorescence revealed that the majority of arrested cells contained a single SPB. Normas development of microtubules between the duplicated SPSs was rarely observed. Multinuclear cells with abnormal microtubule array were detected in small fraction. Taken together with the phenotype of the rlk1 null mutation, these results imply that ROK1 is required for cell cycle progression at the G1/S stage.

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