著者

Ryoko Ando Asako Sakaue-Sawano Keiko Shoda Atsushi Miyawaki

出版者

Japan Society for Cell Biology

雑誌

Cell Structure and Function (ISSN:03867196)

巻号頁・発行日

pp.23028, (Released:2023-06-30)

被引用文献数

7

---

We cloned and characterized two new coral fluorescent proteins: h2-3 and 1-41. h2-3 formed an obligate dimeric complex and exhibited bright green fluorescence. On the other hand, 1-41 formed a highly multimeric complex and exhibited dim red fluorescence. We engineered 1-41 into AzaleaB5, a practically useful red-emitting fluorescent protein for cellular labeling applications. We fused h2-3 and AzaleaB5 to the ubiquitination domains of human Geminin and Cdt1, respectively, to generate a new color variant of Fucci (Fluorescent Ubiquitination-based Cell-Cycle Indicator): Fucci5. We found Fucci5 provided more reliable nuclear labeling for monitoring cell-cycle progression than the 1st and 2nd generations that used mAG/mKO2 and mVenus/mCherry, respectively. Key Words: fluorescent protein, cell cycle, time-lapse imaging, flow cytometry

著者

Ryoko Ando Asako Sakaue-Sawano Keiko Shoda Atsushi Miyawaki

出版者

Japan Society for Cell Biology

雑誌

Cell Structure and Function (ISSN:03867196)

巻号頁・発行日

vol.48, no.2, pp.135-144, 2023 (Released:2023-07-29)

参考文献数

24

被引用文献数

7

---

We cloned and characterized two new coral fluorescent proteins: h2-3 and 1-41. h2-3 formed an obligate dimeric complex and exhibited bright green fluorescence. On the other hand, 1-41 formed a highly multimeric complex and exhibited dim red fluorescence. We engineered 1-41 into AzaleaB5, a practically useful red-emitting fluorescent protein for cellular labeling applications. We fused h2-3 and AzaleaB5 to the ubiquitination domains of human Geminin and Cdt1, respectively, to generate a new color variant of Fucci (Fluorescent Ubiquitination-based Cell-Cycle Indicator): Fucci5. We found Fucci5 provided more reliable nuclear labeling for monitoring cell-cycle progression than the 1st and 2nd generations that used mAG/mKO2 and mVenus/mCherry, respectively.Key words: fluorescent protein, cell cycle, time-lapse imaging, flow cytometry

著者

Hiroyuki Katayama Akitsugu Yamamoto Noboru Mizushima Tamotsu Yoshimori Atsushi Miyawaki

出版者

Japan Society for Cell Biology

雑誌

Cell Structure and Function (ISSN:03867196)

巻号頁・発行日

vol.33, no.1, pp.1-12, 2008 (Released:2008-03-27)

参考文献数

29

被引用文献数

115 192

---

Green fluorescent protein (GFP) from the jellyfish Aequorea victoria, its GFP variants (Aequorea GFPs), and more recently the novel GFP-like proteins from Anthozoa have greatly advanced our technologies for fluorescently labeling cells, organelles, and proteins. It has been shown, however, that some GFP-like proteins have a tendency to oligomerize and aggregate. Transfection of GFP-like proteins into cultured mammalian cells results in bright punctate structures, which are thought to be cytosolic protein aggregates. In this study, we demonstrate that these structures are not cytosolic aggregates but lysosomes that have accumulated the GFP-like proteins. Our biochemical and immunocytochemical experiments have revealed that certain GFP-like proteins expressed in the cytosol enter lysosomes possibly by an autophagy-related mechanism, but retain their fluorescence because of resistance not only to acidity but also to lysosomal proteases.

著者

Atsushi Miyawaki Yasuki Kobayashi Ichiro Kawachi

出版者

Japan Epidemiological Association

雑誌

Journal of Epidemiology (ISSN:09175040)

巻号頁・発行日

vol.30, no.2, pp.67-73, 2020-02-05 (Released:2020-02-05)

参考文献数

49

被引用文献数

12

---

Background: The association of sensory loss with mortality remains unclear. We aimed to explore the associations of hearing loss (HL), visual loss (VL), and dual sensory loss (DSL) with survival.Methods: Data came from the Komo-Ise study cohort in Gunma Prefecture, Japan, where the community-dwelling residents aged 40–69 years were followed up from 1993 to 2010. We analyzed 9,522 individuals who answered the follow-up questionnaires in 2000 (average age 64 [range, 47 to 77] years in 2000). The primary exposures were “HL only,” “VL only,” or “DSL”, with “no HL/VL” as the reference. These sensory loss statuses were assessed by asking the difficulty in hearing conversation or reading newspaper even with aids in the follow-up questionnaires in 2000. All-cause and cause-specific mortality were ascertained from linkage to death certificate data. Cox proportional hazards models adjusting for confounders, including demographic factors, socioeconomic status, and health status, were used. Potential mediators (depression, walking disability, and social participation) were additionally adjusted for.Results: There were 1,105 deaths over the 10-year follow-up. After adjustment for the potential confounders, HL and DSL were associated with increased all-cause mortality (hazard ratios of 1.74 [95% CI, 1.18–2.57] and 1.63 [95% CI, 1.09–2.42], respectively). Potential mediators explained a modest portion of the association. As for cause-specific mortality, HL was associated with increased cancer mortality, while VL and DSL were associated with increased cardiovascular disease mortality.Conclusions: Self-reported HL and DSL may be risk factors of mortality among middle-aged or elderly Japanese populations.