Cargando…
Identification of Novel Mutations in Arabidopsis thaliana DOF 4.2 Coding Gene
Purpose: DOF (DNA-binding with One Finger) proteins are plant-specific transcription factors which mediate numerous biological processes. The purpose of the current study is to report new naturally occurring mutations in the gene encoding for one of the members of DOF proteins named DOF 4.2. Methods...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Tabriz University of Medical Sciences
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8421617/ https://www.ncbi.nlm.nih.gov/pubmed/34513631 http://dx.doi.org/10.34172/apb.2021.064 |
_version_ | 1783749120984350720 |
---|---|
author | Jamshidi Kandjani, Omid Rahbar-Shahrouziasl, Mahdieh Alizadeh, Ali Akbar Hamzeh-Mivehroud, Maryam Dastmalchi, Siavoush |
author_facet | Jamshidi Kandjani, Omid Rahbar-Shahrouziasl, Mahdieh Alizadeh, Ali Akbar Hamzeh-Mivehroud, Maryam Dastmalchi, Siavoush |
author_sort | Jamshidi Kandjani, Omid |
collection | PubMed |
description | Purpose: DOF (DNA-binding with One Finger) proteins are plant-specific transcription factors which mediate numerous biological processes. The purpose of the current study is to report new naturally occurring mutations in the gene encoding for one of the members of DOF proteins named DOF 4.2. Methods: The expression of zinc finger domain of DOF 4.2 (DOF 4.2-ZF) was investigated by first synthesis of cDNA library using different parts of Arabidopsis thaliana plant. Then the coding sequence for zinc finger domain of DOF 4.2 protein was prepared using nested PCR experiment and cloned into pGEX-6P-1 expression vector. Finally, the prepared construct was used for protein expression. Furthermore, molecular dynamics (MD) simulation was carried out to predict DNA binding affinity of DOF 4.2-ZF using AMBER package. Results: For the first time a new variant of DOF 4.2-ZF protein with three mutations was detected. One of the mutations is silent while the other two mutations lead to amino acid replacement (S18G) as well as introduction of a stop codon ultimately resulting in a truncated protein production. In order to investigate whether the truncated form is able to recognize DNA binding motif, MD simulations were carried out and the results showed that the chance of binding of DOF 4.2-ZF protein to cognate DNA in its truncated form is very low. Conclusion: The findings demonstrated that the observed mutations adversely affect the DNA binding ability of the truncated form of DOF4.2 if it is expressed in the mutant variant of A. thaliana used in this study. |
format | Online Article Text |
id | pubmed-8421617 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Tabriz University of Medical Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-84216172021-09-09 Identification of Novel Mutations in Arabidopsis thaliana DOF 4.2 Coding Gene Jamshidi Kandjani, Omid Rahbar-Shahrouziasl, Mahdieh Alizadeh, Ali Akbar Hamzeh-Mivehroud, Maryam Dastmalchi, Siavoush Adv Pharm Bull Research Article Purpose: DOF (DNA-binding with One Finger) proteins are plant-specific transcription factors which mediate numerous biological processes. The purpose of the current study is to report new naturally occurring mutations in the gene encoding for one of the members of DOF proteins named DOF 4.2. Methods: The expression of zinc finger domain of DOF 4.2 (DOF 4.2-ZF) was investigated by first synthesis of cDNA library using different parts of Arabidopsis thaliana plant. Then the coding sequence for zinc finger domain of DOF 4.2 protein was prepared using nested PCR experiment and cloned into pGEX-6P-1 expression vector. Finally, the prepared construct was used for protein expression. Furthermore, molecular dynamics (MD) simulation was carried out to predict DNA binding affinity of DOF 4.2-ZF using AMBER package. Results: For the first time a new variant of DOF 4.2-ZF protein with three mutations was detected. One of the mutations is silent while the other two mutations lead to amino acid replacement (S18G) as well as introduction of a stop codon ultimately resulting in a truncated protein production. In order to investigate whether the truncated form is able to recognize DNA binding motif, MD simulations were carried out and the results showed that the chance of binding of DOF 4.2-ZF protein to cognate DNA in its truncated form is very low. Conclusion: The findings demonstrated that the observed mutations adversely affect the DNA binding ability of the truncated form of DOF4.2 if it is expressed in the mutant variant of A. thaliana used in this study. Tabriz University of Medical Sciences 2021-06 2020-05-09 /pmc/articles/PMC8421617/ /pubmed/34513631 http://dx.doi.org/10.34172/apb.2021.064 Text en ©2021 The Authors. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution (CC BY), which permits unrestricted use, distribution, and reproduction in any medium, as long as the original authors and source are cited. No permission is required from the authors or the publishers. |
spellingShingle | Research Article Jamshidi Kandjani, Omid Rahbar-Shahrouziasl, Mahdieh Alizadeh, Ali Akbar Hamzeh-Mivehroud, Maryam Dastmalchi, Siavoush Identification of Novel Mutations in Arabidopsis thaliana DOF 4.2 Coding Gene |
title |
Identification of Novel Mutations in Arabidopsis thaliana DOF 4.2 Coding Gene
|
title_full |
Identification of Novel Mutations in Arabidopsis thaliana DOF 4.2 Coding Gene
|
title_fullStr |
Identification of Novel Mutations in Arabidopsis thaliana DOF 4.2 Coding Gene
|
title_full_unstemmed |
Identification of Novel Mutations in Arabidopsis thaliana DOF 4.2 Coding Gene
|
title_short |
Identification of Novel Mutations in Arabidopsis thaliana DOF 4.2 Coding Gene
|
title_sort | identification of novel mutations in arabidopsis thaliana dof 4.2 coding gene |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8421617/ https://www.ncbi.nlm.nih.gov/pubmed/34513631 http://dx.doi.org/10.34172/apb.2021.064 |
work_keys_str_mv | AT jamshidikandjaniomid identificationofnovelmutationsinarabidopsisthalianadof42codinggene AT rahbarshahrouziaslmahdieh identificationofnovelmutationsinarabidopsisthalianadof42codinggene AT alizadehaliakbar identificationofnovelmutationsinarabidopsisthalianadof42codinggene AT hamzehmivehroudmaryam identificationofnovelmutationsinarabidopsisthalianadof42codinggene AT dastmalchisiavoush identificationofnovelmutationsinarabidopsisthalianadof42codinggene |