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Conserved Residues in the C-Terminal Domain Affect the Structure and Function of CYP38 in Arabidopsis

Arabidopsis cyclophilin38 (CYP38) is a thylakoid lumen protein critial for PSII assembly and maintenance, and its C-terminal region serves as the target binding domain. We hypothesized that four conserved residues (R290, F294, Q372, and F374) in the C-terminal domain are critical for the structure a...

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Autores principales: Shi, Lujing, Du, Lele, Wen, Jingru, Zong, Xiumei, Zhao, Wene, Wang, Juan, Xu, Min, Wang, Yuhua, Fu, Aigen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7959726/
https://www.ncbi.nlm.nih.gov/pubmed/33732275
http://dx.doi.org/10.3389/fpls.2021.630644
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author Shi, Lujing
Du, Lele
Wen, Jingru
Zong, Xiumei
Zhao, Wene
Wang, Juan
Xu, Min
Wang, Yuhua
Fu, Aigen
author_facet Shi, Lujing
Du, Lele
Wen, Jingru
Zong, Xiumei
Zhao, Wene
Wang, Juan
Xu, Min
Wang, Yuhua
Fu, Aigen
author_sort Shi, Lujing
collection PubMed
description Arabidopsis cyclophilin38 (CYP38) is a thylakoid lumen protein critial for PSII assembly and maintenance, and its C-terminal region serves as the target binding domain. We hypothesized that four conserved residues (R290, F294, Q372, and F374) in the C-terminal domain are critical for the structure and function of CYP38. In yeast two-hybrid and protein pull-down assays, CYP38s with single-sited mutations (R290A, F294A, Q372A, or F374A) did not interact with the CP47 E-loop as the wild-type CYP38. In contrast, CYP38 with the R290A/F294A/Q372A/F374A quadruple mutation could bind the CP47 E-loop. Gene transformation analysis showed that the quadruple mutation prevented CYP38 to efficiently complement the mutant phenotype of cyp38. The C-terminal domain half protein with the quadruple mutation, like the wild-type one, could interact with the N-terminal domain or the CP47 E-loop in vitro. The cyp38 plants expressing CYP38 with the quadruple mutation showed a similar BN-PAGE profile as cyp38, but distinct from the wild type. The CYP38 protein with the quadruple mutation associated with the thylakoid membrane less efficiently than the wild-type CYP38. We concluded that these four conserved residues are indispensable as changes of all these residues together resulted in a subtle conformational change of CYP38 and reduced its intramolecular N-C interaction and the ability to associate with the thylakoid membrane, thus impairing its function in chloroplast.
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spelling pubmed-79597262021-03-16 Conserved Residues in the C-Terminal Domain Affect the Structure and Function of CYP38 in Arabidopsis Shi, Lujing Du, Lele Wen, Jingru Zong, Xiumei Zhao, Wene Wang, Juan Xu, Min Wang, Yuhua Fu, Aigen Front Plant Sci Plant Science Arabidopsis cyclophilin38 (CYP38) is a thylakoid lumen protein critial for PSII assembly and maintenance, and its C-terminal region serves as the target binding domain. We hypothesized that four conserved residues (R290, F294, Q372, and F374) in the C-terminal domain are critical for the structure and function of CYP38. In yeast two-hybrid and protein pull-down assays, CYP38s with single-sited mutations (R290A, F294A, Q372A, or F374A) did not interact with the CP47 E-loop as the wild-type CYP38. In contrast, CYP38 with the R290A/F294A/Q372A/F374A quadruple mutation could bind the CP47 E-loop. Gene transformation analysis showed that the quadruple mutation prevented CYP38 to efficiently complement the mutant phenotype of cyp38. The C-terminal domain half protein with the quadruple mutation, like the wild-type one, could interact with the N-terminal domain or the CP47 E-loop in vitro. The cyp38 plants expressing CYP38 with the quadruple mutation showed a similar BN-PAGE profile as cyp38, but distinct from the wild type. The CYP38 protein with the quadruple mutation associated with the thylakoid membrane less efficiently than the wild-type CYP38. We concluded that these four conserved residues are indispensable as changes of all these residues together resulted in a subtle conformational change of CYP38 and reduced its intramolecular N-C interaction and the ability to associate with the thylakoid membrane, thus impairing its function in chloroplast. Frontiers Media S.A. 2021-02-25 /pmc/articles/PMC7959726/ /pubmed/33732275 http://dx.doi.org/10.3389/fpls.2021.630644 Text en Copyright © 2021 Shi, Du, Wen, Zong, Zhao, Wang, Xu, Wang and Fu. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Shi, Lujing
Du, Lele
Wen, Jingru
Zong, Xiumei
Zhao, Wene
Wang, Juan
Xu, Min
Wang, Yuhua
Fu, Aigen
Conserved Residues in the C-Terminal Domain Affect the Structure and Function of CYP38 in Arabidopsis
title Conserved Residues in the C-Terminal Domain Affect the Structure and Function of CYP38 in Arabidopsis
title_full Conserved Residues in the C-Terminal Domain Affect the Structure and Function of CYP38 in Arabidopsis
title_fullStr Conserved Residues in the C-Terminal Domain Affect the Structure and Function of CYP38 in Arabidopsis
title_full_unstemmed Conserved Residues in the C-Terminal Domain Affect the Structure and Function of CYP38 in Arabidopsis
title_short Conserved Residues in the C-Terminal Domain Affect the Structure and Function of CYP38 in Arabidopsis
title_sort conserved residues in the c-terminal domain affect the structure and function of cyp38 in arabidopsis
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7959726/
https://www.ncbi.nlm.nih.gov/pubmed/33732275
http://dx.doi.org/10.3389/fpls.2021.630644
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