Evaluation of Variation on Myostatin (MSTN) Gene of Turkish Donkey Populations in Thrace Region of Turkey

Öz

Çalışma, Türkiye’de Trakya bölgesinde yetiştirilen 90 eşekte MSTN genindeki varyasyonunu belirlemeyi amaçlamıştır. GDF-8 (growth differentiation factor 8) olarak da adlandırılan miyostatin (MSTN), farklılaşma büyüme faktörü β (TGF-β) süper ailesinin bir parçasıdır ve memeli türlerinde kas kütlesi, büyüme ve gelişme üzerinde negatif düzenleyici rolü bulunmaktadır. MSTN geni, iskelet kası büyümesini olumsuz yönde düzenler ve iskelet kaslarının homeostazında önemli bir role sahiptir. Ayrıca, kas liflerinde protein dengesi Myostatin faktörü tarafından desteklenmektedir. MSTN geninin toplam 866 bç uzunluğunda kısmi intron 1, 2 ve ekzon 2 bölgeleri çoğaltılmış ve PCR ürünleri DNA dizilimi kullanılarak analiz edilmiştir. Bu çalışmada MSTN geninin ikinci ekzon bölgesinde proteinde amino asit değişikliğine neden olmayan yeni bir sinonim SNP g.4183919 G>A belirlenmiştir. G>A değişimi, lösin amino asidinde sessiz bir mutasyona neden olmuştur. Sinonim mutasyonlar nedeniyle mRNA düzeyinde ve proteinin işlevselliğinde değişiklikler meydana gelebilmektedir. Lösin, kas kütlesi kaybını önleyebilen ve miyostatin ekspresyonunu engelleyen önemli bir amino asit olduğundan, bu çalışmada eşeklerde Leu'nun sessiz mutasyonunun eşeklerin kas kütlesini ve fiziksel yapısını değiştirebileceği söylenebilir. Mutant lösin, kas kaybını önlemede daha düşük bir etkiye sahip olduğundan, daha fazla Myostatin protein ekspresyonuna neden olabilmektedir. Türk eşeklerinde daha önce atlarda bulunan bu mutasyonlar tespit edilememiştir, bu da eşeklerin yarış kabiliyeti için daha az gereksinime sahip olduklarını göstermektedir. Tespit edilen SNP ilk olarak ortaya konmuş ve mevcut çalışma ile Türk eşeklerinde MSTN geninin DNA dizileri ilk kez ortaya çıkartılmıştır. MSTN geninin dizileri, MW970078-MW970079 erişim numarasıyla NCBI GenBank'a girilmiştir. Eşek ırklarında MSTN genindeki SNP'lerin ve bunların ekonomik öneme sahip morfolojik karakterlerle ilişkisinin protein ve moleküler düzeyde belirlenmesi amacıyla daha ileri çalışmalara ihtiyaç vardır.

Anahtar Kelimeler

• MSTN
• Eşek
• Trakya
• SNP
• Sekanslama

Evaluation of Variation on Myostatin (MSTN) Gene of Turkish Donkey Populations in Thrace Region of Turkey

Öz

The study aimed to determine the MSTN gene variation in 90 donkeys reared in the Thrace region of Turkey. Myostatin (MSTN), also named GDF-8 (growth differentiation factor 8) is a part of the transforming growth factor β (TGF-β) superfamily and it has a negative regulator role on muscle mass, growth and development in mammalian species. MSTN gene regulates the skeletal muscle growth in a negative way and has a significant role in homeostasis of skeletal muscles. Also, in muscle fibers balance of protein has been promoted by Myostatin factor. The total of 866 bp long partial intron 1 and 2, whole exon 2 regions of MSTN gene was amplified and PCR products analysed using DNA sequencing. In this study, a novel synonymous SNP was determined as g.4183919 G>A in the second exon region of the MSTN gene which does not cause an amino acid change in the protein. The G>A transition caused a silent mutation in leucine (leu) amino acid. Alterations in mRNA level and functionality of protein can occur due to synonymous mutations. Since leucine is an important amino acid that can avoid muscle mass loss and inhibits the expression of Myostatin, it can be said that silent mutation of Leu in donkeys may have altered the muscle mass and physical factor of donkeys in this study. Mutant leucine may have a lower efficient effect on preventing loss of muscles and causes more Myostatin protein expression. The identified SNP was firstly released and the DNA sequences of the MSTN gene in Turkish donkeys was revealed for the first time with recent study. Turkish donkeys lacked these mutations that were identified before in horses, which cause for the less might require for race ability of donkeys. The sequences of MSTN gene were submitted to the NCBI GenBank with the accession number: MW970078- MW970079. Further studies are needed to conduct, on protein and molecular levels, SNPs on the MSTN gene and their association with the morphological characters that may affect economic traits in donkey breeds.

Anahtar Kelimeler

• MSTN
• Donkey
• Thrace
• SNP
• Sequencing

Kaynakça

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