Mapping the membrane orientation of auxin homeostasis
regulators PIN5 and PIN8 in Arabidopsis thaliana root cells
reveals their divergent topology

J 2024

Mapping the membrane orientation of auxin homeostasis regulators PIN5 and PIN8 in Arabidopsis thaliana root cells reveals their divergent topology

SEIFU, Yewubnesh Wendimu; Vendula PUKYŠOVÁ; Nikola RÝDZA; Veronika BILANOVIČOVÁ; Marta ZWIEWKA et. al.

Základní údaje

Originální název

Mapping the membrane orientation of auxin homeostasis regulators PIN5 and PIN8 in Arabidopsis thaliana root cells reveals their divergent topology

Autoři

SEIFU, Yewubnesh Wendimu (231 Etiopie, domácí); Vendula PUKYŠOVÁ (203 Česká republika, domácí); Nikola RÝDZA (703 Slovensko, domácí); Veronika BILANOVIČOVÁ (703 Slovensko, domácí); Marta ZWIEWKA (616 Polsko, domácí); Marek SEDLÁČEK (203 Česká republika, domácí) a Tomasz NODZYNSKI (616 Polsko, domácí)

Vydání

Plant Methods, BMC, 2024, 1746-4811

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10611 Plant sciences, botany

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 4.400

Kód RIV

RIV/00216224:14740/24:00137718

Organizační jednotka

Středoevropský technologický institut

DOI

https://doi.org/10.1186/s13007-024-01182-7

UT WoS

001237623400001

EID Scopus

2-s2.0-85195414498

Klíčová slova anglicky

Arabidopsis thaliana; Auxin homeostasis and transport; Digitonin PM-permeabilizing protocols; Endoplasmic reticulum; Immunocytochemistry; Membrane topology determination methods; PIN auxin efflux carriers

Štítky

CF CELLIM, CF PLANT, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 15. 4. 2025 08:10, Mgr. Eva Dubská

Anotace

V originále

PIN proteins establish the auxin concentration gradient, which coordinates plant growth. PIN1-4 and 7 localized at the plasma membrane (PM) and facilitate polar auxin transport while the endoplasmic reticulum (ER) localized PIN5 and PIN8 maintain the intracellular auxin homeostasis. Although an antagonistic activity of PIN5 and PIN8 proteins in regulating the intracellular auxin homeostasis and other developmental events have been reported, the membrane topology of these proteins, which might be a basis for their antagonistic function, is poorly understood. In this study we optimized digitonin based PM-permeabilizing protocols coupled with immunocytochemistry labeling to map the membrane topology of PIN5 and PIN8 in Arabidopsis thaliana root cells. Our results indicate that, except for the similarities in the orientation of the N-terminus, PIN5 and PIN8 have an opposite orientation of the central hydrophilic loop and the C-terminus, as well as an unequal number of transmembrane domains (TMDs). PIN8 has ten TMDs with groups of five alpha-helices separated by the central hydrophilic loop (HL) residing in the ER lumen, and its N- and C-terminals are positioned in the cytoplasm. However, the topology of PIN5 comprises nine TMDs. Its N-terminal end and the central HL face the cytoplasm while its C-terminus resides in the ER lumen. Overall, this study shows that PIN5 and PIN8 proteins have a divergent membrane topology while introducing a toolkit of methods for studying membrane topology of integral proteins including those localized at the ER membrane.

Návaznosti

GJ20-20860Y, projekt VaV

Název: Odhalení nových regulátorů vnitrobuněčného transportu zapojených do stresové reakce rostlin.

Investor: Grantová agentura ČR, Unravelling novel intracellular trafficking regulators involved in plant stress response.

LQ1601, projekt VaV

Název: CEITEC 2020 (Akronym: CEITEC2020)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CEITEC 2020

90250, velká výzkumná infrastruktura

Název: Czech-BioImaging III

Citovat

SEIFU, Yewubnesh Wendimu; Vendula PUKYŠOVÁ; Nikola RÝDZA; Veronika BILANOVIČOVÁ; Marta ZWIEWKA; Marek SEDLÁČEK a Tomasz NODZYNSKI. Mapping the membrane orientation of auxin homeostasis regulators PIN5 and PIN8 in Arabidopsis thaliana root cells reveals their divergent topology. Plant Methods. BMC, 2024, roč. 20, č. 1, s. 1-16. ISSN 1746-4811. Dostupné z: https://doi.org/10.1186/s13007-024-01182-7.

@article{2453668,
   author = {Seifu, Yewubnesh Wendimu and Pukyšová, Vendula and Rýdza, Nikola and Bilanovičová, Veronika and Zwiewka, Marta and Sedláček, Marek and Nodzynski, Tomasz},
   article_number = {1},
   doi = {https://doi.org/10.1186/s13007-024-01182-7},
   keywords = {Arabidopsis thaliana; Auxin homeostasis and transport; Digitonin PM-permeabilizing protocols; Endoplasmic reticulum; Immunocytochemistry; Membrane topology determination methods; PIN auxin efflux carriers},
   language = {eng},
   issn = {1746-4811},
   journal = {Plant Methods},
   title = {Mapping the membrane orientation of auxin homeostasis regulators PIN5 and PIN8 in Arabidopsis thaliana root cells reveals their divergent topology},
   url = {https://plantmethods.biomedcentral.com/articles/10.1186/s13007-024-01182-7},
   volume = {20},
   year = {2024}
}

TY  - JOUR
ID  - 2453668
AU  - Seifu, Yewubnesh Wendimu - Pukyšová, Vendula - Rýdza, Nikola - Bilanovičová, Veronika - Zwiewka, Marta - Sedláček, Marek - Nodzynski, Tomasz
PY  - 2024
TI  - Mapping the membrane orientation of auxin homeostasis regulators PIN5 and PIN8 in Arabidopsis thaliana root cells reveals their divergent topology
JF  - Plant Methods
VL  - 20
IS  - 1
SP  - 1-16
EP  - 1-16
PB  - BMC
SN  - 17464811
KW  - Arabidopsis thaliana
KW  - Auxin homeostasis and transport
KW  - Digitonin PM-permeabilizing protocols
KW  - Endoplasmic reticulum
KW  - Immunocytochemistry
KW  - Membrane topology determination methods
KW  - PIN auxin efflux carriers
UR  - https://plantmethods.biomedcentral.com/articles/10.1186/s13007-024-01182-7
N2  - PIN proteins establish the auxin concentration gradient, which coordinates plant growth. PIN1-4 and 7 localized at the plasma membrane (PM) and facilitate polar auxin transport while the endoplasmic reticulum (ER) localized PIN5 and PIN8 maintain the intracellular auxin homeostasis. Although an antagonistic activity of PIN5 and PIN8 proteins in regulating the intracellular auxin homeostasis and other developmental events have been reported, the membrane topology of these proteins, which might be a basis for their antagonistic function, is poorly understood. In this study we optimized digitonin based PM-permeabilizing protocols coupled with immunocytochemistry labeling to map the membrane topology of PIN5 and PIN8 in Arabidopsis thaliana root cells. Our results indicate that, except for the similarities in the orientation of the N-terminus, PIN5 and PIN8 have an opposite orientation of the central hydrophilic loop and the C-terminus, as well as an unequal number of transmembrane domains (TMDs). PIN8 has ten TMDs with groups of five alpha-helices separated by the central hydrophilic loop (HL) residing in the ER lumen, and its N- and C-terminals are positioned in the cytoplasm. However, the topology of PIN5 comprises nine TMDs. Its N-terminal end and the central HL face the cytoplasm while its C-terminus resides in the ER lumen. Overall, this study shows that PIN5 and PIN8 proteins have a divergent membrane topology while introducing a toolkit of methods for studying membrane topology of integral proteins including those localized at the ER membrane.
ER  -

SEIFU, Yewubnesh Wendimu; Vendula PUKYŠOVÁ; Nikola RÝDZA; Veronika BILANOVIČOVÁ; Marta ZWIEWKA; Marek SEDLÁČEK a Tomasz NODZYNSKI. Mapping the membrane orientation of auxin homeostasis regulators PIN5 and PIN8 in Arabidopsis thaliana root cells reveals their divergent topology. \textit{Plant Methods}. BMC, 2024, roč.~20, č.~1, s.~1-16. ISSN~1746-4811. Dostupné z: https://doi.org/10.1186/s13007-024-01182-7.

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