Expression of Human PTEN-L in a Yeast Heterologous Model Unveils Specific N-Terminal Motifs Controlling PTEN-L Subcellular Localization and Function

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Fernández Acero, Teresa and Bertalmio, Eleonora and Luna, Sandra and Mingo, Janire and Bravo Plaza, Ignacio and Rodríguez Escudero, Isabel and Molina Martín, María and Pulido, Rafael and Jiménez Cid, Víctor (2019) Expression of Human PTEN-L in a Yeast Heterologous Model Unveils Specific N-Terminal Motifs Controlling PTEN-L Subcellular Localization and Function. Cells, 8 (12). p. 1512. ISSN 2073-4409

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Official URL: https://doi.org/10.3390/cells8121512




Abstract

The tumour suppressor PTEN is frequently downregulated, mutated or lost in several types of tumours and congenital disorders including PHTS (PTEN Hamartoma Tumour Syndrome) and ASD (Autism Spectrum Disorder). PTEN is a lipid phosphatase whose activity over the lipid messenger PIP3 counteracts the stimulation of the oncogenic phosphatidylinositol 3-kinase (PI3K) pathway. Recently, several extended versions of PTEN produced in the cell by alternative translation initiation have been described, among which, PTEN-L and PTEN-M represent the longest isoforms. We previously developed a humanized yeast model in which the expression of PI3K in Saccharomyces cerevisiae led to growth inhibition that could be suppressed by co-expression of PTEN. Here, we show that the expression of PTEN-L and PTEN-M in yeast results in robust counteracting of PI3K-dependent growth inhibition. N-terminally tagged GFP-PTEN-L was sharply localized at the yeast plasma membrane. Point mutations of a putative membrane-binding helix located at the PTEN-L extension or its deletion shifted localization to nuclear. Also, a shift from plasma membrane to nucleus was observed in mutants at basic amino acid clusters at the PIP2-binding motif, and at the Cα2 and CBR3 loops at the C2 domain. In contrast, C-terminally tagged PTEN-L-GFP displayed mitochondrial localization in yeast, which was shifted to plasma membrane by removing the first 22 PTEN-L residues. Our results suggest an important role of the N-terminal extension of alternative PTEN isoforms on their spatial and functional regulation.


Item Type:Article
Uncontrolled Keywords:PTEN; phosphoinositides; subcellular localization; alternative translation initiation; Saccharomyces cerevisiae; PI3K; heterologous expression
Subjects:Medical sciences > Pharmacy > Microbiology
Medical sciences > Pharmacy > Parasitology
ID Code:67103
Deposited On:28 Jul 2021 13:42
Last Modified:19 Aug 2021 07:13

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