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Rab5(Q79L) interacts with the carboxyl terminus of RUFY3

International Journal of Biological Sciences
Ivyspring International Publisher
Publication Date
  • Letter To The Editor
  • Biology


Int. J. Biol. Sci. 2010, 6 187 IInntteerrnnaattiioonnaall JJoouurrnnaall ooff BBiioollooggiiccaall SScciieenncceess 2010; 6(2):187-189 © Ivyspring International Publisher. All rights reserved Letter to the editor Rab5(Q79L) interacts with the carboxyl terminus of RUFY3 Hitomi Yoshida1, Naoko Okumura1, Yasuko Kitagishi1, Naoki Shirafuji2, Satoru Matsuda1 1. Department of Environmental Health, Nara Women's University, Nara 630-8506, Japan. 2. Department of Hematology/Oncology, Teikyo University School of Medicine, Tokyo, Japan. Corresponding author: Satoru Matsuda, Email: [email protected] Received: 2010.02.04; Accepted: 2010.03.31; Published: 2010.04.04 Key words: atherosclerosis; RUFY3; Rab5; RUN domain; protein interaction The small GTPase Rab5 controls the fusogenic properties of early endosomes through activation of effector proteins. Expression of a GTPase-defective mutant, Rab5(Q79L), is known to cause formation of enlarged early endosomes. To identify proteins inte- racting with this GTP-bound form of active Rab5 [1], we used a two-hybrid system to screen a human fetal cDNA library with a human Rab5A protein bearing a mutation of Q79L as bait. The positive clone cDNAs were reconfirmed to be interaction-positive by using an independent yeast clone containing the bait. Then, more than 3 positive preys were identified, and sub- sequent sequence analysis revealed one of the preys to be RUFY3: RUN and FYVE domain containing 3. We performed double immunefluorescent staining for human RUFY3 and human Rab5A(Q79L) in 3Y1 cells. Analysis with confocal microscopy clearly showed that the Rab5(Q79L) and the RUFY3 colocalized in large vesicle structures in 3Y1 cells (data not shown). This indicated an interaction between RUFY3 and Rab5(Q79L), which may be altered by the nucleo- tide-bound state of Rab5. We checked this possibility using other Rab5 mutant. The Rab5(S34N) mutant preferentially binds GDP

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