Martynoside rescues 5-fluorouracil-impaired ribosome biogenesis by stabilizing RPL27A

Mengying Hong; Yushen Du; Dongdong Chen; Yuan Shi; Menglong Hu; Kejun Tang; Zhuping Hong; Xiangzhi Meng; Wan Xu; Gaoqi Wu; Yuanyuan Yao; Liubo Chen; Wenteng Chen; Chit Ying Lau; Li Sheng; Tian-Hao Zhang; Haigen Huang; Zheyu Fang; Yong Shen; Fangfang Sun; Jing Qian; Haibin Qu; Shu Zheng; Suzhan Zhang; Kefeng Ding; Ren Sun

doi:10.1016/j.scib.2023.07.018

Martynoside rescues 5-fluorouracil-impaired ribosome biogenesis by stabilizing RPL27A

Sci Bull (Beijing). 2023 Aug 15;68(15):1662-1677. doi: 10.1016/j.scib.2023.07.018. Epub 2023 Jul 15.

Authors

Mengying Hong¹, Yushen Du², Dongdong Chen³, Yuan Shi⁴, Menglong Hu⁵, Kejun Tang⁶, Zhuping Hong⁷, Xiangzhi Meng⁸, Wan Xu⁹, Gaoqi Wu¹⁰, Yuanyuan Yao¹¹, Liubo Chen⁶, Wenteng Chen¹², Chit Ying Lau⁹, Li Sheng⁴, Tian-Hao Zhang¹³, Haigen Huang⁴, Zheyu Fang⁶, Yong Shen⁶, Fangfang Sun⁶, Jing Qian⁷, Haibin Qu⁷, Shu Zheng¹⁴, Suzhan Zhang¹⁴, Kefeng Ding¹⁴, Ren Sun¹⁵

Affiliations

¹ Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education), The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China; Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles 90095, USA; Zhejiang Province Key Laboratory of Molecular Biology in Medical Sciences, Hangzhou 310009, China; Zhejiang Provincial Clinical Research Center for Cancer, Cancer Center of Zhejiang University, Hangzhou 310009, China.
² Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education), The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China; Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles 90095, USA; Zhejiang Province Key Laboratory of Molecular Biology in Medical Sciences, Hangzhou 310009, China; Zhejiang Provincial Clinical Research Center for Cancer, Cancer Center of Zhejiang University, Hangzhou 310009, China. Electronic address: lilyduyushen@zju.edu.cn.
³ Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education), The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China; Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles 90095, USA.
⁴ Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles 90095, USA.
⁵ Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education), The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China; School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
⁶ Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education), The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China.
⁷ Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
⁸ School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China; Department of Microbiology, Immunology, & Molecular Genetics, University of California, Los Angeles 90095, USA; Center for Infectious Disease Research, School of Life Sciences, Institute for Advanced Studies, Westlake University, Hangzhou 310024, China.
⁹ School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
¹⁰ Institute of Immunology, Zhejiang University School of Medicine, Hangzhou 310058, China.
¹¹ Department of Colorectal Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
¹² College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
¹³ Molecular Biology Institute, University of California, Los Angeles 90095, USA.
¹⁴ Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education), The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China; Zhejiang Province Key Laboratory of Molecular Biology in Medical Sciences, Hangzhou 310009, China; Zhejiang Provincial Clinical Research Center for Cancer, Cancer Center of Zhejiang University, Hangzhou 310009, China.
¹⁵ Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles 90095, USA; School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China; Molecular Biology Institute, University of California, Los Angeles 90095, USA; Center for Infectious Disease Research, School of Life Sciences, Institute for Advanced Studies, Westlake University, Hangzhou 310024, China. Electronic address: sunren@westlake.edu.cn.

PMID: 37481436
DOI: 10.1016/j.scib.2023.07.018

Abstract

Martynoside (MAR), a bioactive component in several well-known tonic traditional Chinese herbs, exhibits pro-hematopoietic activity during 5-fluorouracil (5-FU) treatment. However, the molecular target and the mechanism of MAR are poorly understood. Here, by adopting the mRNA display with a library of even-distribution (md-LED) method, we systematically examined MAR-protein interactions in vitro and identified the ribosomal protein L27a (RPL27A) as a key cellular target of MAR. Structural and mutational analysis confirmed the specific interaction between MAR and the exon 4,5-encoded region of RPL27A. MAR attenuated 5-FU-induced cytotoxicity in bone marrow nucleated cells, increased RPL27A protein stability, and reduced the ubiquitination of RPL27A at lys92 (K92) and lys94 (K94). Disruption of MAR binding at key residues of RPL27A completely abolished the MAR-induced stabilization. Furthermore, by integrating label-free quantitative ubiquitination proteomics, transcriptomics, and ribosome function assays, we revealed that MAR restored RPL27A protein levels and thus rescued ribosome biogenesis impaired by 5-FU. Specifically, MAR increased mature ribosomal RNA (rRNA) abundance, prevented ribosomal protein degradation, facilitated ribosome assembly, and maintained nucleolar integrity. Collectively, our findings characterize the target of a component of Chinese medicine, reveal the importance of ribosome biogenesis in hematopoiesis, and open up a new direction for improving hematopoiesis by targeting RPL27A.

Keywords: Martynoside; RPL27A; Ribosome biogenesis; mRNA display.

MeSH terms

Biological Assay*
Bone Marrow Cells
Caffeine
Fluorouracil* / pharmacology

Substances

martynoside
Fluorouracil
Caffeine