The second intracellular loop of the yeast Trk1 potassium transporter is involved in regulation of activity, and interaction with 14–3-3 proteins

Masaryk, Jakub; Kale, Deepika; Pohl, Pavel; Ruiz-Castilla, F. J.; Zimmermannová, Olga; Obšilová, Veronika; Ramos, J.; Sychrová, Hana

doi:https://dx.doi.org/10.1016/j.csbj.2023.04.019

Number of the records: 1

The second intracellular loop of the yeast Trk1 potassium transporter is involved in regulation of activity, and interaction with 14–3-3 proteins

1.

SYSNO ASEP	0571383
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	The second intracellular loop of the yeast Trk1 potassium transporter is involved in regulation of activity, and interaction with 14–3-3 proteins
Author(s)	Masaryk, Jakub (FGU-C) Kale, Deepika (FGU-C) Pohl, Pavel (FGU-C)_ORCID Ruiz-Castilla, F. J. (ES) Zimmermannová, Olga (FGU-C)_{RID, ORCID} Obšilová, Veronika (FGU-C)_{RID, ORCID, SAI} Ramos, J. (ES) Sychrová, Hana (FGU-C)_{RID, ORCID}
Source Title	Computational and Structural Biotechnology Journal. - : Elsevier - ISSN 2001-0370 Roč. 21, April (2023), s. 2705-2716
Number of pages	12 s.
Language	eng - English
Country	NL - Netherlands
Keywords	potassium ion uptake ; Saccharomyces cerevisiae ; phosphorylation ; Trk1 ; 14–3–3 proteins
OECD category	Microbiology
R&D Projects	GA20-04420S GA ČR - Czech Science Foundation (CSF)
	GA21-08985S GA ČR - Czech Science Foundation (CSF)
	LTC20005 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
Method of publishing	Open access
Institutional support	FGU-C - RVO:67985823
UT WOS	000989417300001
EID SCOPUS	85153376257
DOI	10.1016/j.csbj.2023.04.019
Annotation	Potassium is an essential intracellular ion, and a sufficient intracellular concentration of it is crucial for many processes, therefore it is fundamental for cells to precisely regulate K+ uptake and efflux through the plasma membrane. The uniporter Trk1 is a key player in K+ acquisition in yeasts. The TRK1 gene is expressed at a low and stable level, thus the activity of the transporter needs to be regulated at a posttranslational level. S. cerevisiae Trk1 changes its activity and affinity for potassium ion quickly and according to both internal and external concentrations of K+, as well as the membrane potential. The molecular basis of these changes has not been elucidated, though phosphorylation is thought to play an important role. In this study, we examined the role of the second, short, and highly conserved intracellular hydrophilic loop of Trk1 (IL2), and identified two phosphorylable residues (Ser882 and Thr900) as very important for 1) the structure of the loop and consequently for the targeting of Trk1 to the plasma membrane, and 2) the upregulation of the transporter’s activity reaching maximal affinity under low external K+ conditions. Moreover, we identified three residues (Thr155, Ser414, and Thr900) within the Trk1 protein as strong candidates for interaction with 14–3–3 regulatory proteins, and showed, in an in vitro experiment, that phosphorylated Thr900 of the IL2 indeed binds to both isoforms of yeast 14–3–3 proteins, Bmh1 and Bmh2.
Workplace	Institute of Physiology
Contact	Lucie Trajhanová, lucie.trajhanova@fgu.cas.cz, Tel.: 241 062 400
Year of Publishing	2024
Electronic address	https://doi.org/10.1016/j.csbj.2023.04.019

Number of the records: 1