The main objective of this study was to analyze the physiological activities of Scd6: its localization, interactions, functions and regulatory mechanisms in RNA decay pathway. By investigating Scd6 interacting factors, I found that Scd6 interacts with Hmt1. In addition, I examined the roles of Scd6 for P-body formation and its interactions with other decapping activators.
First, by two-hybrid screening assay using Scd6 as bait, I found Scd6 interacting candidates including known Scd6-interacting partner, Dcp1. In this study, I focused on analyzing the interaction with Hmt1, a mono- and asymmetric di-arginine methyltransferase. I found that Hmt1 directly binds to Scd6. Deletion of HMT1 results in loss of Scd6 methylation in the RGG motifs.
This Hmt1-dependent methylation was involved in Scd6 subcellular localization under glucose starvation. In addition, I revealed that Scd6 plays a role with Dhh1 and Edc3. Scd6 also gave synthetic impact on P-body formation with Dhh1.
Although Scd6 is highly methylated in physiological condition that is required
for its efficient targeting to P-bodies, this methylation is not required for Scd6
function on cell growth at 25
oC or overexpression-mediated induction of P-body
formation. These results show that Scd6 accumulated to facilitate the P-body
formation and arginine residues within the RGG motif were not strictly required
for cell growth in dhh1 mutation background, at least under physiological condition.
I show herein the regulation of Scd6 physiological activities by
Hmt1-dependent arginine methylation. The present study provides a new insight
into how post-translational modifications of mRNP components could regulate
the gene expression in response to external conditions.
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ACKNOWLEDGEMENT
I would like to express my sincere gratitude to my advisor, Dr. Kenji Irie in Department of Molecular Cell Biology, University of Tsukuba, for accepting me as a graduate student in his laboratory and offering me kind guidance and resources for the present study. I am also thankful to Dr. Yasuyuki Suda in Department of Molecular Cell Biology, University of Tsukuba, for his kind direct instruction and support throughout the course of this research. This dissertation would not have been possible without their encouragement and support.
I am so grateful to Dr. Keiichi Izumikawa in Global Innovation Research Organization, Tokyo University of Agriculture and Technology, for his support to my research, particularly mass spectrometry analysis. I am also thankful to Ms.
Kaoru Irie and Mr. Kei Muroi in Department of Molecular Cell Biology,
University of Tsukuba for their significant support and contribution to my genetic
analyses.
ドキュメント内
Hmt1との相互作用とメチル化によるScd6の生理学的機能の解析
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