A histone modifier, ASXL1, interacts with NONO and is involved in paraspeckle formation in hematopoietic cells

Keita Yamamoto; Susumu Goyama; Shuhei Asada; Takeshi Fujino; Taishi Yonezawa; Naru Sato; Reina Takeda; Akiho Tsuchiya; Tomofusa Fukuyama; Yosuke Tanaka; Akihiko Yokoyama; Hikaru Toya; Ayana Kon; Yasuhito Nannya; Rena Onoguchi-Mizutani; Shinichi Nakagawa; Tetsuro Hirose; Seishi Ogawa; Nobuyoshi Akimitsu; Toshio Kitamura

doi:10.1016/j.celrep.2021.109576

A histone modifier, ASXL1, interacts with NONO and is involved in paraspeckle formation in hematopoietic cells

Cell Rep. 2021 Aug 24;36(8):109576. doi: 10.1016/j.celrep.2021.109576.

Authors

Keita Yamamoto¹, Susumu Goyama¹, Shuhei Asada², Takeshi Fujino¹, Taishi Yonezawa¹, Naru Sato¹, Reina Takeda¹, Akiho Tsuchiya¹, Tomofusa Fukuyama¹, Yosuke Tanaka¹, Akihiko Yokoyama³, Hikaru Toya⁴, Ayana Kon⁵, Yasuhito Nannya⁵, Rena Onoguchi-Mizutani⁶, Shinichi Nakagawa⁴, Tetsuro Hirose⁷, Seishi Ogawa⁵, Nobuyoshi Akimitsu⁶, Toshio Kitamura⁸

Affiliations

¹ Division of Cellular Therapy, Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
² Division of Cellular Therapy, Institute of Medical Science, The University of Tokyo, Tokyo, Japan; The Institute of Laboratory Animals, Tokyo Women's Medical University, Tokyo, Japan.
³ National Cancer Center Tsuruoka Metabolomics Laboratory, Yamagata, Japan.
⁴ RNA Biology Laboratory, Faculty of Pharmaceutical Sciences, Hokkaido University, Hokkaido, Japan.
⁵ Department of Pathology and Tumor Biology, Kyoto University, Kyoto, Japan.
⁶ Isotope Science Center, The University of Tokyo, Tokyo, Japan.
⁷ Graduate School of Frontier Biosciences, Osaka University, Suita, Japan.
⁸ Division of Cellular Therapy, Institute of Medical Science, The University of Tokyo, Tokyo, Japan. Electronic address: [email protected].

PMID: 34433054
DOI: 10.1016/j.celrep.2021.109576

Abstract

Paraspeckles are membraneless organelles formed through liquid-liquid phase separation and consist of multiple proteins and RNAs, including NONO, SFPQ, and NEAT1. The role of paraspeckles and the component NONO in hematopoiesis remains unknown. In this study, we show histone modifier ASXL1 is involved in paraspeckle formation. ASXL1 forms phase-separated droplets, upregulates NEAT1 expression, and increases NONO-NEAT1 interactions through the C-terminal intrinsically disordered region (IDR). In contrast, a pathogenic ASXL mutant (ASXL1-MT) lacking IDR does not support the interaction of paraspeckle components. Furthermore, paraspeckles are disrupted and Nono localization is abnormal in the cytoplasm of hematopoietic stem and progenitor cells (HSPCs) derived from ASXL1-MT knockin mice. Nono depletion and the forced expression of cytoplasmic NONO impair the repopulating potential of HSPCs, as does ASXL1-MT. Our study indicates a link between ASXL1 and paraspeckle components in the maintenance of normal hematopoiesis.

Keywords: ASXL1; HSC; LLPS; MDS; clonal hematopoiesis; hematopoiesis; phase separation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism*
Female
HL-60 Cells
HeLa Cells
Hematopoiesis
Hematopoietic Stem Cells / metabolism*
Humans
Mice
Mice, Transgenic
Paraspeckles / genetics
Paraspeckles / metabolism*
RNA-Binding Proteins / genetics
RNA-Binding Proteins / metabolism*
Repressor Proteins / genetics
Repressor Proteins / metabolism*
THP-1 Cells

Substances

ASXL1 protein, human
Asxl1 protein, mouse
DNA-Binding Proteins
NONO protein, human
Nono protein, mouse
RNA-Binding Proteins
Repressor Proteins