Structure and mechanism of an actin-dependent bacterial phosphoryl AMPylase

Tao-Tao Chen; Qiuhua Lu; Si-Ru Zheng; Jiaqi Fu; Jing Chen; Lina Kang; Juhong Wu; Jiwei Luo; Jiangyang Tong; Siying Li; Xiangliang Li; Shan Li; Jinyu Li; Shaoyuan Wang; Yue Feng; Zhao-Qing Luo; Songying Ouyang

doi:10.1038/s41589-025-01945-w

Structure and mechanism of an actin-dependent bacterial phosphoryl AMPylase

Nat Chem Biol. 2025 Jun 30. doi: 10.1038/s41589-025-01945-w. Online ahead of print.

Authors

Tao-Tao Chen^#¹, Qiuhua Lu^#¹, Si-Ru Zheng^#¹, Jiaqi Fu^#², Jing Chen^#³, Lina Kang¹, Juhong Wu⁴, Jiwei Luo⁵, Jiangyang Tong¹, Siying Li², Xiangliang Li¹, Shan Li⁵, Jinyu Li⁴, Shaoyuan Wang³, Yue Feng⁶, Zhao-Qing Luo⁷, Songying Ouyang⁸

Affiliations

¹ Key Laboratory of Microbial Pathogenesis and Interventions of Fujian Province University, Key Laboratory of Innate Immune Biology of Fujian Province, Biomedical Research Center of South China, College of Life Sciences, Fujian Normal University, Fuzhou, China.
² Department of Respiratory Medicine, Center for Infectious Diseases and Pathogen Biology, Key Laboratory of Organ Regeneration and Transplantation of the Ministry of Education, State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, The First Hospital of Jilin University, Changchun, China.
³ Department of Hematology, Fujian Institute of Hematology, Fujian Provincial Key Laboratory on Hematology, Fujian Medical University Union Hospital, Fuzhou, China.
⁴ College of Chemistry, Fuzhou University, Fuzhou, China.
⁵ National Key Laboratory of Agricultural Microbiology, College of Biomedicine and Health, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, China.
⁶ Beijing Advanced Innovation Center for Soft Matter Science and Engineering, Beijing Key Laboratory of Bioprocess, State Key Laboratory of Chemical Resource Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, China.
⁷ Department of Biological Sciences, Purdue University, West Lafayette, IN, USA. luoz@purdue.edu.
⁸ Key Laboratory of Microbial Pathogenesis and Interventions of Fujian Province University, Key Laboratory of Innate Immune Biology of Fujian Province, Biomedical Research Center of South China, College of Life Sciences, Fujian Normal University, Fuzhou, China. ouyangsy@fjnu.edu.cn.

^# Contributed equally.

PMID: 40588486
DOI: 10.1038/s41589-025-01945-w

Abstract

The two effectors LnaB and MavL of Legionella pneumophila coordinate the conversion of phosphoribosyl ubiquitin (PR-Ub) released by reversal of ubiquitination induced by members of the SidE effector family into functional Ub. LnaB acts as an actin-dependent phosphoryl AMPylase that converts PR-Ub into ADP-ribosylated (ADPR)-Ub. Catalysis by LnaB requires the conserved SHE motif present in a large family of bacterial toxins. Here we describe a series of structures of LnaB in complex with the cofactor actin and the substrate PR-Ub and ATP. LnaB harbors both adenylyltransferase and ATPase activities, which reveal an adenylylation mechanism involved in a two-step catalytic process. Actin performs a unique activation mechanism that promotes the recruitment of PR-Ub by LnaB to activate LnaB's ATPase activity through interacting with LnaB and PR-Ub. Mechanisms derived from this series of structures covering the process of LnaB action establish an important biochemical basis for protein AMPylation.

Abstract

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