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The tobacco chloroplast YCF4 gene is essential for transcriptional gene regulation and plants photoautotrophic growth

A tobacco chloroplast hypothetical open reading frame 4 (YCF4) has been reported as a non-essential assembly factor for photosynthesis based on an incomplete knockout of YCF4, just 93 of 184 amino acids from the N-terminus were knocked out. On the other hand, we removed the complete sequence of YCF4...

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Autores principales: Khan, Muhammad Sarwar, Riaz, Rimsha, Majid, Muhammad, Mehmood, Kashif, Mustafa, Ghulam, Joyia, Faiz Ahmad
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9638951/
https://www.ncbi.nlm.nih.gov/pubmed/36352880
http://dx.doi.org/10.3389/fpls.2022.1014236
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author Khan, Muhammad Sarwar
Riaz, Rimsha
Majid, Muhammad
Mehmood, Kashif
Mustafa, Ghulam
Joyia, Faiz Ahmad
author_facet Khan, Muhammad Sarwar
Riaz, Rimsha
Majid, Muhammad
Mehmood, Kashif
Mustafa, Ghulam
Joyia, Faiz Ahmad
author_sort Khan, Muhammad Sarwar
collection PubMed
description A tobacco chloroplast hypothetical open reading frame 4 (YCF4) has been reported as a non-essential assembly factor for photosynthesis based on an incomplete knockout of YCF4, just 93 of 184 amino acids from the N-terminus were knocked out. On the other hand, we removed the complete sequence of YCF4 from tobacco chloroplasts and observed that ΔYCF4 plants were unable to survive photoautotrophically as their growth was hampered in the absence of an external carbon supply, clearly showing that the YCF4 is essential for photosynthesis. Initially, the aadA gene was introduced into the tobacco plastome replacing the complete YCF4 gene through homologous recombination events. The replacement of YCF4 with aadA was confirmed by PCR and Southern blot analysis in ΔYCF4 plants. Homoplasmic ΔYCF4 plants had a light green phenotype, and the leaves became pale yellow as the plants grew older. The structure of chloroplasts of ΔYCF4 mutants of light green phenotype was studied using a transmission electron microscope (TEM), and the micrographs demonstrated structural anomalies in the chloroplasts; including shape, size, and grana stacking compared to the wild-type plants. Further, transcriptome analysis revealed that the expression of PSI, PSII, and ribosomal genes remained unchanged in ∆YCF4 plants. On the other hand, transcriptome levels of rbcL (Ribulose 1,5-bisphosphate carboxylase/oxygenase large subunit), LHC (Light-Harvesting Complex), and ATP Synthase (atpB and atpL) decreased, indicating that the YCF4 has the function(s) in addition to assembling the photosynthetic complex. This was confirmed by in-silico protein-protein interactions of full-length YCF4 as well as 93 and 91 of 184 amino acids from N- and C-termini of the full-length protein, which revealed that the C-terminus (91 aa) of YCF4 is important in interacting with other chloroplast proteins. These findings provide genetic support for the plastid YCF4 gene’s critical role in regulating the plastid gene expression and assembling the photosynthetic complex.
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spelling pubmed-96389512022-11-08 The tobacco chloroplast YCF4 gene is essential for transcriptional gene regulation and plants photoautotrophic growth Khan, Muhammad Sarwar Riaz, Rimsha Majid, Muhammad Mehmood, Kashif Mustafa, Ghulam Joyia, Faiz Ahmad Front Plant Sci Plant Science A tobacco chloroplast hypothetical open reading frame 4 (YCF4) has been reported as a non-essential assembly factor for photosynthesis based on an incomplete knockout of YCF4, just 93 of 184 amino acids from the N-terminus were knocked out. On the other hand, we removed the complete sequence of YCF4 from tobacco chloroplasts and observed that ΔYCF4 plants were unable to survive photoautotrophically as their growth was hampered in the absence of an external carbon supply, clearly showing that the YCF4 is essential for photosynthesis. Initially, the aadA gene was introduced into the tobacco plastome replacing the complete YCF4 gene through homologous recombination events. The replacement of YCF4 with aadA was confirmed by PCR and Southern blot analysis in ΔYCF4 plants. Homoplasmic ΔYCF4 plants had a light green phenotype, and the leaves became pale yellow as the plants grew older. The structure of chloroplasts of ΔYCF4 mutants of light green phenotype was studied using a transmission electron microscope (TEM), and the micrographs demonstrated structural anomalies in the chloroplasts; including shape, size, and grana stacking compared to the wild-type plants. Further, transcriptome analysis revealed that the expression of PSI, PSII, and ribosomal genes remained unchanged in ∆YCF4 plants. On the other hand, transcriptome levels of rbcL (Ribulose 1,5-bisphosphate carboxylase/oxygenase large subunit), LHC (Light-Harvesting Complex), and ATP Synthase (atpB and atpL) decreased, indicating that the YCF4 has the function(s) in addition to assembling the photosynthetic complex. This was confirmed by in-silico protein-protein interactions of full-length YCF4 as well as 93 and 91 of 184 amino acids from N- and C-termini of the full-length protein, which revealed that the C-terminus (91 aa) of YCF4 is important in interacting with other chloroplast proteins. These findings provide genetic support for the plastid YCF4 gene’s critical role in regulating the plastid gene expression and assembling the photosynthetic complex. Frontiers Media S.A. 2022-10-24 /pmc/articles/PMC9638951/ /pubmed/36352880 http://dx.doi.org/10.3389/fpls.2022.1014236 Text en Copyright © 2022 Khan, Riaz, Majid, Mehmood, Mustafa and Joyia https://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
Khan, Muhammad Sarwar
Riaz, Rimsha
Majid, Muhammad
Mehmood, Kashif
Mustafa, Ghulam
Joyia, Faiz Ahmad
The tobacco chloroplast YCF4 gene is essential for transcriptional gene regulation and plants photoautotrophic growth
title The tobacco chloroplast YCF4 gene is essential for transcriptional gene regulation and plants photoautotrophic growth
title_full The tobacco chloroplast YCF4 gene is essential for transcriptional gene regulation and plants photoautotrophic growth
title_fullStr The tobacco chloroplast YCF4 gene is essential for transcriptional gene regulation and plants photoautotrophic growth
title_full_unstemmed The tobacco chloroplast YCF4 gene is essential for transcriptional gene regulation and plants photoautotrophic growth
title_short The tobacco chloroplast YCF4 gene is essential for transcriptional gene regulation and plants photoautotrophic growth
title_sort tobacco chloroplast ycf4 gene is essential for transcriptional gene regulation and plants photoautotrophic growth
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9638951/
https://www.ncbi.nlm.nih.gov/pubmed/36352880
http://dx.doi.org/10.3389/fpls.2022.1014236
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