Dataset Information

Stk38 interactomics by proximity ligation

ABSTRACT: STK38 (aka NDR1) is a Hippo pathway serine/threonine protein kinase with multifarious functions in normal and cancer cells. Using a context-dependent proximity labelling assay, we discovered that STK38 modulates its interaction with more than 250 proteins depending on the context. Upon starvation-induced autophagy, STK38 associates with cytoplasmic proteins while upon ECM detachment it binds to mainly nuclear proteins. This differential subcellular-localization-dependent activity is caused by the XPO1 (Exportin-1, CRM1) dependent nuclear/cytoplasmic shuttling of STK38. STK38 associates with nuclear related partners upon ECM detachment and with cytoplasmic related ones upon autophagy, exhibiting a nuclear/cytoplasmic shuttling under the dependency of XPO1 (Exportin-1, aka CRM1). We further uncovered that the XPO1-mediated export of STK38 is dependent on phosphorylation of XPO1s serin residue 1055 by STK38 itself. In addition to regulating its own nuclear export, STK38 also controls the subcellular distribution of Beclin1, a key regulator of autophagy. Moreover, the regulation of XPO1 by STK38 mediates the nuclear exclusion of YAP1. Collectively, our results reveal that functions of STK38 are linked to the XPO1-mediated subcellular distribution of STK38 and key regulators. These observations show that unrelated cellular functions can be regulated by a same mechanism controlled by a single kinase and demonstrate a novel mechanism of XPO1-dependent cargo export regulation y phosphorylation of XPO1s C-terminal auto-inhibitory domain.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

SUBMITTER: Chiara guerrera

LAB HEAD: I Chiara GUERRERA

PROVIDER: PXD011968 | Pride | 2019-09-26

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20161027_JaCO_1_1_DR_4340.raw	Raw
	20161027_JaCO_1_2_DR_4341.raw	Raw
	20161027_JaCO_1_3_DR_4342.raw	Raw
	20161027_JaCO_3_1_DR_4346.raw	Raw
	20161027_JaCO_3_2_DR_4347.raw	Raw

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Publications

STK38 kinase acts as XPO1 gatekeeper regulating the nuclear export of autophagy proteins and other cargoes.

Martin Alexandre Pj AP Jacquemyn Maarten M Lipecka Joanna J Chhuon Cerina C Aushev Vasily N VN Meunier Brigitte B Singh Manish K MK Carpi Nicolas N Piel Matthieu M Codogno Patrice P Hergovich Alexander A Parrini Maria Carla MC Zalcman Gerard G Guerrera Ida Chiara IC Daelemans Dirk D Camonis Jacques H JH

EMBO reports 20190923 11

STK38 (also known as NDR1) is a Hippo pathway serine/threonine protein kinase with multifarious functions in normal and cancer cells. Using a context-dependent proximity-labeling assay, we identify more than 250 partners of STK38 and find that STK38 modulates its partnership depending on the cellular context by increasing its association with cytoplasmic proteins upon nutrient starvation-induced autophagy and with nuclear ones during ECM detachment. We show that STK38 shuttles between the nucleu ...[more]

PMID: 31544310

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Project description:SF3B1 mutations are the most frequent spliceosomal alterations across cancers, yet no successful therapy exists to target this pathway. Previous findings from a phase 2 clinical trial of the XPO1 inhibitor selinexor in patients with high-risk myelodysplastic neoplasms (MDS) relapsed or refractory to hypomethylating agents (HMA) revealed increased activity in patients with SF3B1 mutations. XPO1 (Exportin-1) is responsible for the export of over 200 proteins, but also plays a role in the transport of multiple RNA species, including small nuclear RNAs (snRNAs), ribosomal RNAs (rRNAs), and select messenger RNAs (mRNAs) out of the nucleus. We therefore hypothesized that XPO1 inhibition perturbs RNA export and may preferentially affect SF3B1 mutants via altered splicing, given the role of XPO1 in exporting snRNAs, which form the catalytic portion of the spliceosome. To evaluate the mechanism underlying preferentially sensitivity of SF3B1-mutants to XPO1 inhibition, we performed nuclear and cytoplasmic fraction followed by RNA sequencing before and after XPO1 inhibition in SF3B1 wildtype and SF3B1 K666N cells (subcellular RNA-seq). Whole transcriptomic analysis of subcellular RNA-seq data revealed more nuclear retention of global RNA transcripts after XPO1 inhibition in the SF3B1 mutant cell line. Similarly, we performed subcellular RNA-seq for small RNAs and found snRNAs to be increased in the nucleus after XPO1 inhibition in the SF3B1 mutant cells. We then performed total cellular RNA sequencing to understand the effect of XPO1 inhibition on global RNA expression and RNA splicing. Differential gene expression analysis identified that XPO1 inhibition had the greatest effect on cell cycle in SF3B1 wildtype cells but in SF3B1-mutant cells differentiation pathways were more significantly affected. Alternative splicing analysis showed increased 3’ alternative splicing events in SF3B1 mutant after XPO1 inhibition. These results signify the mechanistic basis for preferential sensitivity of SF3B1 mutant cells to nuclear export inhibition arises through nuclear retention of spliceosomal snRNAs and select mRNAs that result in perturbation of differentiation pathways.