5-methylcytosine modification by Plasmodium NSUN2 stabilizes mRNA and mediates the development of gametocytes

Meng Liu; Gangqiang Guo; Pengge Qian; Jianbing Mu; Binbin Lu; Xiaoqin He; Yanting Fan; Xiaomin Shang; Guang Yang; Shijun Shen; Wenju Liu; Liping Wang; Liang Gu; Quankai Mu; Xinyu Yu; Yuemeng Zhao; Richard Culleton; Jun Cao; Lubin Jiang; Thomas E Wellems; Jing Yuan; Cizhong Jiang; Qingfeng Zhang

doi:10.1073/pnas.2110713119

5-methylcytosine modification by Plasmodium NSUN2 stabilizes mRNA and mediates the development of gametocytes

Proc Natl Acad Sci U S A. 2022 Mar 1;119(9):e2110713119. doi: 10.1073/pnas.2110713119.

Authors

Meng Liu^{1

2}, Gangqiang Guo^{1

2

3}, Pengge Qian⁴, Jianbing Mu⁵, Binbin Lu³, Xiaoqin He⁶, Yanting Fan³, Xiaomin Shang³, Guang Yang^{1

2}, Shijun Shen^{1

2}, Wenju Liu^{1

2}, Liping Wang^{1

2}, Liang Gu^{1

2}, Quankai Mu⁷, Xinyu Yu⁶, Yuemeng Zhao^{3

8}, Richard Culleton^{9

10}, Jun Cao⁶, Lubin Jiang⁸, Thomas E Wellems¹¹, Jing Yuan¹², Cizhong Jiang^{13

2}, Qingfeng Zhang¹⁴

Affiliations

¹ Key Laboratory of Spine and Spinal Cord Injury Repair and Regeneration of Ministry of Education, Orthopaedic Department of Tongji Hospital, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
² Shanghai Key Laboratory of Signaling and Disease Research, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
³ Key Laboratory of Spine and Spinal Cord Injury Repair and Regeneration of Ministry of Education, Tongji Hospital, Clinical Center for Brain and Spinal Cord Research, School of Medicine, Tongji University, Shanghai 200092, China.
⁴ State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Signaling Network, School of Life Sciences, Xiamen University, Fujian 361005, China.
⁵ Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD 20892-8132.
⁶ National Health Commission Key Laboratory of Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi 214064, China.
⁷ Department of Immunogenetics, Institute of Tropical Medicine, Nagasaki University, Nagasaki 852-8523, Japan.
⁸ Unit of Human Parasite Molecular and Cell Biology, Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
⁹ Division of Molecular Parasitology, Proteo-Science Centre, Ehime University, Ehime 790-8577, Japan.
¹⁰ Department of Protozoology, Institute of Tropical Medicine, Nagasaki University, Nagasaki 852-8521, Japan.
¹¹ Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD 20892-8132; twellems@niaid.nih.gov yuanjing@xmu.edu.cn czjiang@tongji.edu.cn qfzhang@tongji.edu.cn.
¹² State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Signaling Network, School of Life Sciences, Xiamen University, Fujian 361005, China; twellems@niaid.nih.gov yuanjing@xmu.edu.cn czjiang@tongji.edu.cn qfzhang@tongji.edu.cn.
¹³ Key Laboratory of Spine and Spinal Cord Injury Repair and Regeneration of Ministry of Education, Orthopaedic Department of Tongji Hospital, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China; twellems@niaid.nih.gov yuanjing@xmu.edu.cn czjiang@tongji.edu.cn qfzhang@tongji.edu.cn.
¹⁴ Key Laboratory of Spine and Spinal Cord Injury Repair and Regeneration of Ministry of Education, Tongji Hospital, Clinical Center for Brain and Spinal Cord Research, School of Medicine, Tongji University, Shanghai 200092, China; twellems@niaid.nih.gov yuanjing@xmu.edu.cn czjiang@tongji.edu.cn qfzhang@tongji.edu.cn.

Abstract

5-methylcytosine (m⁵C) is an important epitranscriptomic modification involved in messenger RNA (mRNA) stability and translation efficiency in various biological processes. However, it remains unclear if m⁵C modification contributes to the dynamic regulation of the transcriptome during the developmental cycles of Plasmodium parasites. Here, we characterize the landscape of m⁵C mRNA modifications at single nucleotide resolution in the asexual replication stages and gametocyte sexual stages of rodent (Plasmodium yoelii) and human (Plasmodium falciparum) malaria parasites. While different representations of m⁵C-modified mRNAs are associated with the different stages, the abundance of the m⁵C marker is strikingly enhanced in the transcriptomes of gametocytes. Our results show that m⁵C modifications confer stability to the Plasmodium transcripts and that a Plasmodium ortholog of NSUN2 is a major mRNA m⁵C methyltransferase in malaria parasites. Upon knockout of P. yoelii nsun2 (pynsun2), marked reductions of m⁵C modification were observed in a panel of gametocytogenesis-associated transcripts. These reductions correlated with impaired gametocyte production in the knockout rodent malaria parasites. Restoration of the nsun2 gene in the knockout parasites rescued the gametocyte production phenotype as well as m⁵C modification of the gametocytogenesis-associated transcripts. Together with the mRNA m⁵C profiles for two species of Plasmodium, our findings demonstrate a major role for NSUN2-mediated m⁵C modifications in mRNA transcript stability and sexual differentiation in malaria parasites.

Keywords: RNA bisulfite sequencing; RNA methyltransferase; epitranscriptomic modifications; gametocytogenesis; gene knockout.

MeSH terms

5-Methylcytosine / chemistry*
Germ Cells
Plasmodium falciparum / genetics
Plasmodium falciparum / growth & development
Plasmodium falciparum / metabolism*
Plasmodium yoelii / genetics
Plasmodium yoelii / growth & development*
Plasmodium yoelii / metabolism*
Protozoan Proteins / metabolism*
RNA, Messenger / metabolism*
Transcriptome

Substances

Protozoan Proteins
RNA, Messenger
5-Methylcytosine

Grants and funding

ZIA AI000483/ImNIH/Intramural NIH HHS/United States