Arabidopsis'te Dawdle geninin FHA alanını ve C-terminal bölgesini kapsayan nokta mutasyonlarının fenotipik karakterizasyonu

Yıl 2024, , 61 - 71, 27.06.2024

Seyit Yüzüak , David Chevalier

https://doi.org/10.29048/makufebed.1482242

Öz

Arabidopsis thaliana'daki gen fonksiyon kaybı alellerinin analizi, DAWDLE (DDL) adı verilen At3G20550 geninde bir mutasyonu ortaya çıkarmış ve DDL genindeki bu mutasyonun pleiotropik fenotiplere neden olduğu ve birçok mikro RNA seviyesini de düşürdüğü rapor edilmiştir. DAWDLE geni, prokaryot ve ökaryotlarda önemli hücresel fonksiyonlara sahip birçok proteinde de bulunan Çatal Başlı-İlişkili Bölge (FHA) alanına sahip bir proteini kodlar. Ancak, DDL proteinin FHA alanının ve C-terminal bölgesinin bu genin işlevi için gerekli olup olmadığı tam olarak bilinmemektedir. Bu çalışmada, DDL geninin FHA alanı ve C-terminal bölgesini kapsayan nokta mutasyonlarının fenotipik karakterizasyonu yapılarak her iki bölgenin DDL fonksiyonunda rolü olup olmadığı analiz edilmiştir. Arabidopsis'in Columbia erecta-105 yabanıl tipinde ‘Genomda Hedefli İndüklenmiş Lokal Lezyonlar’ (Tilling) hat taraması yapılarak DDL geninin FHA ve C-terminal bölgesini kapsayan nokta mutasyonları tespit edilmiş, ardından bu nokta mutasyonları taşıyan homozigot mutantlar belirlenerek bitki boyu, hipokotil ve kök uzunluğu ve tohum sayısı seviyesinde fenotipik olarak karakterize edilmiştir. Mutantların fenotipik analizi, farklı bitki organlarında değişen derecelerde benzer ddl fenotiplerini ortaya çıkarmıştır. Tiller mutant hatlarının tohum sayısındaki düşüş, kök, hipokotil ve gövde uzunluklarındaki kısalma, Arabidopsis'te FHA ve C-terminal bölgesinin DDL geninin fonksiyonu için önemli olabileceğini ortaya koymuştur.
Anahtar Kelimeler: Dawdle, Çatal Başlı-İlişkili Bölge, Genomda Hedefli İndüklenmiş Lokal Lezyonlar, Etil Metan Sülfonat, Arabidopsis

Anahtar Kelimeler

Dawdle, Çatal Başlı İlişkili Bölge, Genomda Hedeflş İndüklenmiş Lokal Lezyonlar, Etil Metan Sülfonat, Arabidopsis

Destekleyen Kurum

Mississippi Eyalet Üniversitesi Biyolojik Bilimler Bölümü

Teşekkür

Araştırmanın yürütülmesi sırasında verdikleri danışmanlık desteği için Dr. Lakshmi Narayanan ve Dr. Ercan Selçuk Ünlü'ye özel teşekkürlerimizi sunarız. Ayrıca, sorumlu yazara Amerika Birleşik Devletleri'nde lisansüstü eğitim alması için finansal destek sağlayan Türkiye Cumhuriyeti Milli Eğitim Bakanlığı'na ve araştırma fonu için Mississippi Eyalet Üniversitesi Biyolojik Bilimler Bölümü'ne minnettarız.

Phenotypic characterization of point mutations spanning FHA domain and C-terminal region of Dawdle gene in Arabidopsis

Öz

The screening analysis of loss-of-function alleles in Arabidopsis thaliana revealed a mutation in the At3G20550 gene, called DAWDLE (DDL). The mutation in the DDL gene causes pleiotropic phenotypes and reduced the levels of several microRNAs. The DAWDLE gene encodes a protein with a Fork Head-Associated (FHA) domain, found in large range of proteins with significant cellular processes in prokaryotes and eukaryotes. However, it is not completely known whether the FHA domain and C-terminal region of the DDL are necessary for its function. The aim of this study was to determine the function of both regions by conducting a phenotypic analysis of point mutations spanning the FHA domain and C-terminal region in DDL Targeted Induced Local Lesions IN Genome (Tilling) screen was performed in the Columbia erecta-105 background of Arabidopsis resulting in point mutations spanning both regions of DDL. The mutants were phenotypically characterized. Height of the plant, hypocotyl and root length, and fertility were measured. Phenotypic analyses of the mutants revealed ddl phenotypes of varying degrees in different organs. Reduction in fertility and shortening in root, hypocotyl and stem lengths of the Tiller mutant lines suggest that the FHA domain and C-terminal region may require for DDL function in Arabidopsis.
Key words: Dawdle, Fork Head-Associated Domain, Targeted Induced Local Lesions in Genome, Ethyl Methane Sulfonate, Arabidopsis

Anahtar Kelimeler

Dawdle, Fork Head-Associated Domain, Targeted Induced Local Lesions in Genome, Ethyl Methane Sulfonate, Arabidopsis

Destekleyen Kurum

Department of Biological Sciences, Mississippi State University

Teşekkür

We would like to give our special thanks to Dr. Lakshmi Narayanan and Dr. Ercan Selcuk Unlu for their advisory support during the conducting of the research. We are also grateful to the Ministry of National Education, The Republic of Turkey, for providing a financial support to the corresponding author for receiving the graduate degrees in the United States, and to the Department of Biological Sciences, Mississippi State University for research funding

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