An integrative synthetic biology approach to interrogating cellular ubiquitin and UFM signaling

Chuanyin Li, Tianting Han, Rong Guo, Peng Chen, Chao Peng, Gali Prag, Ronggui Hu*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Global identification of substrates for PTMs (post-translational modifications) represents a critical but yet dauntingly challenging task in understanding biology and disease pathology. Here we presented a synthetic biology approach, namely ‘YESS’, which coupled Y2H (yeast two hybrid) interactome screening with PTMs reactions reconstituted in bacteria for substrates identification and validation, followed by the functional validation in mammalian cells. Specifically, the sequence-independent Gateway®cloning technique was adopted to afford simultaneous transfer of multiple hit ORFs (open reading frames) between the YESS sub-systems. In proof-of-evidence applications of YESS, novel substrates were identified for UBE3A and UFL1, the E3 ligases for ubiquitination and ufmylation, respectively. Therefore, the YESS approach could serve as a potentially powerful tool to study cellular signaling mediated by different PTMs.

Original languageEnglish
Article number4231
Pages (from-to)1-16
Number of pages16
JournalInternational Journal of Molecular Sciences
Volume21
Issue number12
DOIs
StatePublished - 2 Jun 2020

Funding

FundersFunder number
Aaron Ciechanover of Technion-Israel Institute of Technology and Hai Rao of U.T. San Antonia
Ministry of Science and Technology of China2018YFA0508200, 2018ZX10101004, 2019YFA0802103
National Natural Science Foundation of China81525019, 31800700, 31900804
Chinese Academy of SciencesXDA12040323, XDB19000000
China Postdoctoral Science Foundation2018M642109
Science and Technology Commission of Shanghai Municipality2017SHZDZX01, 19140903500
Israel Science Foundation651/16

    Keywords

    • Post-translational modifications
    • UBE3A
    • UFL1
    • Ubiquitination
    • Ufmylation

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