An N-Terminal Extension to UBA5 Adenylation Domain Boosts UFM1 Activation: Isoform-Specific Differences in Ubiquitin-like Protein Activation

Nadine Soudah, Prasanth Padala, Fouad Hassouna, Manoj Kumar, Bayan Mashahreh, Andrey A. Lebedev, Michail N. Isupov, Einav Cohen-Kfir, Reuven Wiener*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

Modification of proteins by the ubiquitin-like protein, UFM1, requires activation of UFM1 by the E1-activating enzyme, UBA5. In humans, UBA5 possesses two isoforms, each comprising an adenylation domain, but only one containing an N-terminal extension. Currently, the role of the N-terminal extension in UFM1 activation is not clear. Here we provide structural and biochemical data on UBA5 N-terminal extension to understand its contribution to UFM1 activation. The crystal structures of the UBA5 long isoform bound to ATP with and without UFM1 show that the N-terminus not only is directly involved in ATP binding but also affects how the adenylation domain interacts with ATP. Surprisingly, in the presence of the N-terminus, UBA5 no longer retains the 1:2 ratio of ATP to UBA5, but rather this becomes a 1:1 ratio. Accordingly, the N-terminus significantly increases the affinity of ATP to UBA5. Finally, the N-terminus, although not directly involved in the E2 binding, stimulates transfer of UFM1 from UBA5 to the E2, UFC1.

Original languageAmerican English
Pages (from-to)463-478
Number of pages16
JournalJournal of Molecular Biology
Volume431
Issue number3
DOIs
StatePublished - 1 Feb 2019

Bibliographical note

Funding Information:
We thank the staff of beamlines ID29 and ID30A-1 of the European Synchrotron Radiation Facility. This work was supported by, the Marie Curie Career Integration Grant ( PCIG13-GA-2013-630755 ), the Israel Science Foundation (Grant 1383/17 ) and the Israeli Cancer Association (Grant 20180069 ).

Funding Information:
We thank the staff of beamlines ID29 and ID30A-1 of the European Synchrotron Radiation Facility. This work was supported by, the Marie Curie Career Integration Grant (PCIG13-GA-2013-630755), the Israel Science Foundation (Grant 1383/17) and the Israeli Cancer Association (Grant 20180069).

Publisher Copyright:
© 2018 Elsevier Ltd

Keywords

  • E1 activating enzymes
  • UBA5
  • UFM1
  • crystal structure
  • ubiquitin-like proteins

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