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THEMIS is a novel substrate and allosteric activator of SHP1, playing dual roles during T cell development

ABSTRACT: By combining strategies of substrate trapping for PTPs and pupylation-based interaction tagging (PUP-IT) for protein-protein interaction, we develop a systematic proximity labeling methodology PEPSI in identifying novel substrate for PTPs. We apply this approach to SHP1 and surprisingly, identified THEMIS as an uncharacterized substrate for the phosphatase.

ORGANISM(S): Homo Sapiens

SUBMITTER: Gaofeng Fan

PROVIDER: PXD028689 | iProX | Thu Aug 31 00:00:00 BST 2023

REPOSITORIES: iProX

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THEMIS is a substrate and allosteric activator of SHP1, playing dual roles during T cell development.

Zhang Jiali J Jiang Zhenzhou Z Zhang Xueyuan X Yang Ziqun Z Wang Jinjiao J Chen Jialing J Chen Li L Song Minfang M Zhang Yanchun Y Huang Mei M Chen Shengmiao S Xiong Xuexue X Wang Yuetong Y Hao Piliang P Horng Tiffany T Zhuang Min M Zhang Liye L Zuo Erwei E Bai Fang F Zheng Jie J Wang Haopeng H Fan Gaofeng G

Nature structural & molecular biology 20240104 1

THEMIS plays an indispensable role in T cells, but its mechanism of action has remained highly controversial. Using the systematic proximity labeling methodology PEPSI, we identify THEMIS as an uncharacterized substrate for the phosphatase SHP1. Saturated mutagenesis assays and mass spectrometry analysis reveal that phosphorylation of THEMIS at the evolutionally conserved Tyr34 residue is oppositely regulated by SHP1 and the kinase LCK. Similar to THEMIS<sup>-</sup><sup>/-</sup> mice, THEMIS<sup ...[more]

PMID: 38177672

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Project description:Pupylation is a posttranslational protein modification in bacteria resembling ubiquitination in eukaryotes. The prokaryotic ubiquitin-like protein Pup is covalently attached to other proteins via an isopeptide bond between its carboxyterminal glutamate residue and a lysine residue in the target. In mycobacteria, pupylation was shown to mark proteins for unfolding by the ATPase Mpa and subsequent degradation by the proteasome. However, the occurrence of pupylation in species without a proteasome like Corynebacterium glutamicum suggests that degradation may not be the only fate of pupylated proteins. The Îpup mutant senses a stronger iron limitation than the wild type. Among the 125 genes showing at least 2-fold changes in transcript levels in the Îpup mutant (p-value â¤ 0.05) were 54% of all genes known to be regulated by the master regulator of iron homeostasis DtxR (Brune et al., 2006; Wennerhold and Bott, 2006). Except for ftn, which is activated by DtxR and showed a 2-fold decreased mRNA level, the other DtxR target genes showed increased mRNA levels in the Îpup strain. These included ripA, encoding a transcriptional regulator of iron proteins, which represses a number of prominent iron-containing proteins under iron limitation, such as aconitase or succinate dehydrogenase (Wennerhold et al., 2005). 79% of the known RipA target genes showed decreased mRNA levels in the Îpup strain. DNA microarray analyses were performed to compare the mRNA levels of the C. glutamicum Îpup mutant and its parent wild type under iron-limited conditions. The two strains precultivated in CGXII medium with 4% (w/v) glucose and 1 ÂµM FeSO4 were inoculated into fresh medium to an OD600 of 1, cultured for 2 h, and harvested on ice by centrifugation (5 min at 4,000 g and 4Â°C). Please note that the GPL16989 array design comprises oligos for 4 different bacterial genomes. In the GPR-files in this study, all IDs/Names (oligonucleotides) which are not from the host C. glutamicum were replaced by the text EMPTY since only C. glutamicum expression was analyzed.

Dataset Information

THEMIS is a novel substrate and allosteric activator of SHP1, playing dual roles during T cell development

Publications

THEMIS is a substrate and allosteric activator of SHP1, playing dual roles during T cell development.

Similar Datasets

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