@article{65075e400dca45bb9793cf6f43442876,
title = "Structural and Biochemical Study of the Mono-ADP-Ribosyltransferase Domain of SdeA, a Ubiquitylating/Deubiquitylating Enzyme from Legionella pneumophila",
abstract = "Conventional ubiquitylation occurs through an ATP-dependent three-enzyme cascade (E1, E2, and E3) that mediates the covalent conjugation of the C-terminus of ubiquitin to a lysine on the substrate. SdeA, which belongs to the SidE effector family of Legionella pneumophila, can transfer ubiquitin to endoplasmic reticulum-associated Rab-family GTPases in a manner independent of E1 and E2 enzymes. The novel ubiquitin-modifying enzyme SdeA utilizes NAD+ as a cofactor to attach ubiquitin to a serine residue of the substrate. Here, to elucidate the coupled enzymatic reaction of NAD + hydrolysis and ADP-ribosylation of ubiquitin in SdeA, we characterized the mono-ADP-ribosyltransferase domain of SdeA and show that it consists of two sub-domains termed mART-N and mART-C. The crystal structure of the mART-C domain of SdeA was also determined in free form and in complex with NAD+ at high resolution. Furthermore, the spatial orientations of the N-terminal deubiquitylase, phosphodiesterase, mono-ADP-ribosyltransferase, and C-terminal coiled-coil domains within the 180-kDa full-length SdeA were determined. These results provide insight into the unusual ubiquitylation mechanism of SdeA and expand our knowledge on the structure–function of mono-ADP-ribosyltransferases.",
keywords = "Legionella pneumophila, NAD, SdeA, mART, ubiquitin",
author = "Leehyeon Kim and Kwon, {Do Hoon} and Kim, {Bong Heon} and Jiyeon Kim and Park, {Mi Rae} and Park, {Zee Yong} and Song, {Hyun Kyu}",
note = "Funding Information: We thank the staff at beamline 5C, Pohang Accelerator Laboratory, Republic of Korea, and beamline BL-17A, Photon Factory, Japan, for their help with the X-ray data collection. This work was in part performed under the International Collaborative Research Program of Institute for Protein Research, Osaka University (ICR-17-05). Diffraction data were collected at the Osaka University beamline BL44XU at SPring-8 (Harima, Japan; Proposal Nos. 2017A6775 and 2017B6775). We also thank the staff at beamline BL-10C, Photon Factory, Japan, for their help with the SAXS data collection. L.K. is a recipient of a POSCO Science Fellowship of POSCO TJ Park Foundation. This research was supported by a grant from the Samsung Science & Technology Foundation (SSTF-BA 1701-14). Funding Information: We thank the staff at beamline 5C, Pohang Accelerator Laboratory, Republic of Korea, and beamline BL-17A, Photon Factory, Japan, for their help with the X-ray data collection. This work was in part performed under the International Collaborative Research Program of Institute for Protein Research, Osaka University (ICR-17-05). Diffraction data were collected at the Osaka University beamline BL44XU at SPring-8 (Harima, Japan; Proposal Nos. 2017A6775 and 2017B6775). We also thank the staff at beamline BL-10C, Photon Factory, Japan, for their help with the SAXS data collection. L.K. is a recipient of a POSCO Science Fellowship of POSCO TJ Park Foundation. This research was supported by a grant from the Samsung Science & Technology Foundation ( SSTF-BA 1701-14 ). Publisher Copyright: {\textcopyright} 2018 Elsevier Ltd",
year = "2018",
month = aug,
day = "17",
doi = "10.1016/j.jmb.2018.05.043",
language = "English",
volume = "430",
pages = "2843--2856",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "17",
}