Functional analysis of the transcriptional repressor HapX involved in iron homeostasis in filamentous fungi

2015 Fiscal Year Final Research Report

• Project/Area Number: 25450116
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: Meijo University
• Principal Investigator: Kato Masashi 名城大学, 農学部, 教授 (70242849)
• Co-Investigator(Kenkyū-buntansha): SHIMIZU MOTOYUKI 名城大学, 農学部, 助教 (20423535)
• Co-Investigator(Renkei-kenkyūsha): KOBAYASHI TETSUO 名古屋大学, 生命農学研究科, 教授 (20170334) ; TAKAYA NAOKI 筑波大学, 生命環境科学研究科, 教授 (50282322)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: HapX / 鉄の恒常性維持 / ホメオスタシス / CCAAT配列 / Hap複合体 / 転写抑制 / 転写因子 / 転写調節

Outline of Final Research Achievements

(1) Results of EMSA（Electrophoresis Mobility Shift Assay） with some HapX deletion mutants strongly suggested that the 17 amino acid residues in the N-terminal domain plays an important role in the interaction of HapX and HapB/C/E complex. (2) The results of the in vivo analysis showed that the 17 amino acid residues is required for the function of HapX. (3) We have succeeded in obtaining sufficient amount of the recombinant C-terminal domain containing the Cysteine-rich regions. The results of spectroscopic analysis suggested the existence of iron-sulfur clusters. (4)　We established the expression system for a HapX protein with TAP (Tandem Affinity Purification) tag in Aspergillus nidulans. When the HapX was purified under the mild conditions, some proteins interacting with HapX were detected.

Free Research Field

応用微生物学