UBL3 Interacts with PolyQ-Expanded Huntingtin Fragments and Modifies Their Intracellular Sorting

Background/Objectives: UBL3 (Ubiquitin-like 3) is a protein that plays a crucial role in post-translational modifications, particularly in regulating protein transport within small extracellular vesicles. While previous research has predominantly focused on its interactions with α-synuclein, this st...

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Main Authors: Soho Oyama, Hengsen Zhang, Rafia Ferdous, Yuna Tomochika, Bin Chen, Shuyun Jiang, Md. Shoriful Islam, Md. Mahmudul Hasan, Qing Zhai, A. S. M. Waliullah, Yashuang Ping, Jing Yan, Mst. Afsana Mimi, Chi Zhang, Shuhei Aramaki, Yusuke Takanashi, Tomoaki Kahyo, Yoshio Hashizume, Daita Kaneda, Mitsutoshi Setou
Format: Article
Language:English
Published: MDPI AG 2024-10-01
Series:Neurology International
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Online Access:https://www.mdpi.com/2035-8377/16/6/89
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author Soho Oyama
Hengsen Zhang
Rafia Ferdous
Yuna Tomochika
Bin Chen
Shuyun Jiang
Md. Shoriful Islam
Md. Mahmudul Hasan
Qing Zhai
A. S. M. Waliullah
Yashuang Ping
Jing Yan
Mst. Afsana Mimi
Chi Zhang
Shuhei Aramaki
Yusuke Takanashi
Tomoaki Kahyo
Yoshio Hashizume
Daita Kaneda
Mitsutoshi Setou
author_facet Soho Oyama
Hengsen Zhang
Rafia Ferdous
Yuna Tomochika
Bin Chen
Shuyun Jiang
Md. Shoriful Islam
Md. Mahmudul Hasan
Qing Zhai
A. S. M. Waliullah
Yashuang Ping
Jing Yan
Mst. Afsana Mimi
Chi Zhang
Shuhei Aramaki
Yusuke Takanashi
Tomoaki Kahyo
Yoshio Hashizume
Daita Kaneda
Mitsutoshi Setou
author_sort Soho Oyama
collection DOAJ
description Background/Objectives: UBL3 (Ubiquitin-like 3) is a protein that plays a crucial role in post-translational modifications, particularly in regulating protein transport within small extracellular vesicles. While previous research has predominantly focused on its interactions with α-synuclein, this study investigates UBL3’s role in Huntington’s disease (HD). HD is characterized by movement disorders and cognitive impairments, with its pathogenesis linked to toxic, polyglutamine (polyQ)-expanded mutant huntingtin fragments (mHTT). However, the mechanisms underlying the interaction between UBL3 and mHTT remain poorly understood. Methods: To elucidate this relationship, we performed hematoxylin and eosin (HE) staining and immunohistochemistry (IHC) on postmortem brain tissue from HD patients. Gaussia princeps-based split-luciferase complementation assay and co-immunoprecipitation were employed to confirm the interaction between UBL3 and mHTT. Additionally, we conducted a HiBiT lytic detection assay to assess the influence of UBL3 on the intracellular sorting of mHTT. Finally, immunocytochemical staining was utilized to validate the colocalization and distribution of these proteins. Results: Our findings revealed UBL3-positive inclusions in the cytoplasm and nuclei of neurons throughout the striatum of HD patients. We discovered that UBL3 colocalizes and interacts with mHTT and modulates its intracellular sorting. Conclusions: These results suggest that UBL3 may play a significant role in the interaction and sorting of mHTT, contributing to the understanding of its potential implications in the pathophysiology of Huntington’s disease.
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spelling doaj-art-37c8760c4d01449282e2dd4cf521d3442025-08-20T02:01:15ZengMDPI AGNeurology International2035-83772024-10-011661175118810.3390/neurolint16060089UBL3 Interacts with PolyQ-Expanded Huntingtin Fragments and Modifies Their Intracellular SortingSoho Oyama0Hengsen Zhang1Rafia Ferdous2Yuna Tomochika3Bin Chen4Shuyun Jiang5Md. Shoriful Islam6Md. Mahmudul Hasan7Qing Zhai8A. S. M. Waliullah9Yashuang Ping10Jing Yan11Mst. Afsana Mimi12Chi Zhang13Shuhei Aramaki14Yusuke Takanashi15Tomoaki Kahyo16Yoshio Hashizume17Daita Kaneda18Mitsutoshi Setou19Department of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanFirst Department of Surgery, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanChoju Medical Institute, Fukushimura Hospital, Yamanaka-19-14 Noyoricho, Toyohashi 441-8124, Aichi, JapanChoju Medical Institute, Fukushimura Hospital, Yamanaka-19-14 Noyoricho, Toyohashi 441-8124, Aichi, JapanDepartment of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Chuo-ku, Hamamatsu 431-3192, Shizuoka, JapanBackground/Objectives: UBL3 (Ubiquitin-like 3) is a protein that plays a crucial role in post-translational modifications, particularly in regulating protein transport within small extracellular vesicles. While previous research has predominantly focused on its interactions with α-synuclein, this study investigates UBL3’s role in Huntington’s disease (HD). HD is characterized by movement disorders and cognitive impairments, with its pathogenesis linked to toxic, polyglutamine (polyQ)-expanded mutant huntingtin fragments (mHTT). However, the mechanisms underlying the interaction between UBL3 and mHTT remain poorly understood. Methods: To elucidate this relationship, we performed hematoxylin and eosin (HE) staining and immunohistochemistry (IHC) on postmortem brain tissue from HD patients. Gaussia princeps-based split-luciferase complementation assay and co-immunoprecipitation were employed to confirm the interaction between UBL3 and mHTT. Additionally, we conducted a HiBiT lytic detection assay to assess the influence of UBL3 on the intracellular sorting of mHTT. Finally, immunocytochemical staining was utilized to validate the colocalization and distribution of these proteins. Results: Our findings revealed UBL3-positive inclusions in the cytoplasm and nuclei of neurons throughout the striatum of HD patients. We discovered that UBL3 colocalizes and interacts with mHTT and modulates its intracellular sorting. Conclusions: These results suggest that UBL3 may play a significant role in the interaction and sorting of mHTT, contributing to the understanding of its potential implications in the pathophysiology of Huntington’s disease.https://www.mdpi.com/2035-8377/16/6/89ubiquitin-like 3huntington’s diseasepolyglutaminehuntingtin proteininteractionsorting
spellingShingle Soho Oyama
Hengsen Zhang
Rafia Ferdous
Yuna Tomochika
Bin Chen
Shuyun Jiang
Md. Shoriful Islam
Md. Mahmudul Hasan
Qing Zhai
A. S. M. Waliullah
Yashuang Ping
Jing Yan
Mst. Afsana Mimi
Chi Zhang
Shuhei Aramaki
Yusuke Takanashi
Tomoaki Kahyo
Yoshio Hashizume
Daita Kaneda
Mitsutoshi Setou
UBL3 Interacts with PolyQ-Expanded Huntingtin Fragments and Modifies Their Intracellular Sorting
Neurology International
ubiquitin-like 3
huntington’s disease
polyglutamine
huntingtin protein
interaction
sorting
title UBL3 Interacts with PolyQ-Expanded Huntingtin Fragments and Modifies Their Intracellular Sorting
title_full UBL3 Interacts with PolyQ-Expanded Huntingtin Fragments and Modifies Their Intracellular Sorting
title_fullStr UBL3 Interacts with PolyQ-Expanded Huntingtin Fragments and Modifies Their Intracellular Sorting
title_full_unstemmed UBL3 Interacts with PolyQ-Expanded Huntingtin Fragments and Modifies Their Intracellular Sorting
title_short UBL3 Interacts with PolyQ-Expanded Huntingtin Fragments and Modifies Their Intracellular Sorting
title_sort ubl3 interacts with polyq expanded huntingtin fragments and modifies their intracellular sorting
topic ubiquitin-like 3
huntington’s disease
polyglutamine
huntingtin protein
interaction
sorting
url https://www.mdpi.com/2035-8377/16/6/89
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