Structural and Biochemical Study of the Mono-ADP-Ribosyltransferase Domain of SdeA, a Ubiquitylating/Deubiquitylating Enzyme from Legionella pneumophila

Leehyeon Kim, Do Hoon Kwon, Bong Heon Kim, Jiyeon Kim, Mi Rae Park, Zee Yong Park, Hyun Kyu Song

Research output: Contribution to journalArticle

6 Citations (Scopus)

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.

Original languageEnglish
JournalJournal of Molecular Biology
DOIs
Publication statusAccepted/In press - 2018 Jan 1

Keywords

  • Legionella pneumophila
  • mART
  • NAD
  • SdeA
  • ubiquitin

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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