Investigation of GDF9 and BMP15 Polymorphisms in Mehraban Sheep to Find the Missenses as Impact on Protein
محورهای موضوعی : Camelا. احمدی 1 , ف. افراز 2 , ر. طالبی 3 , ع. فرح‌آور 4 , س.م.ف. وحیدی 5
1 - Department of Animal Science, Faculty of Agriculture, Bu Ali Sina University, Hamedan, Iran
2 - Department of Genomics and Animal, Agricultural Biotechnology Research Institute (ABRI), North branch, Rasht, Iran
3 - Department of Animal Science, Faculty of Agriculture, Bu Ali Sina University, Hamedan, Iran
4 - Department of Animal Science, Faculty of Agriculture, Bu Ali Sina University, Hamedan, Iran
5 - Department of Genomics and Animal, Agricultural Biotechnology Research Institute (ABRI), North branch, Rasht, Iran
کلید واژه: Sheep, <, i>, BMP15<, /i>, , functional interaction, <, i>, GDF9<, /i>, , missense, protein structure,
چکیده مقاله :
Utilization of fecundity genes such as GDF9 and BMP15 can help improve reproductive traits in sheep breeding programme. To evaluate effects of missense mutations on protein function, the polymorphisms of GDF9 and BMP15 genes were screened in twelve mehraban sheep using DNA sequencing, followed by protein structure modeling. Six single nucleotide polymorphism (SNPs) known as FecG mutations (G1-G6), were detected in exons 1 and 2 of GDF9 gene. Mutations of G1 (GDF9 exon 1 g.2118 G>A), G4 (GDF9 exon 2 g.3451 T>C) and G6 (GDF9 exon 2 g.3974 G>A) have shown amino acid substitution. None polymorphism was detected in exon 1 and exon 2 of BMP15 gene. Based on identified polymorphisms, individuals were classified into three haplotypes of wild haplotype (without mutation), haplotype A (simultaneous mutations of G1, G2, G3 and G4) and haplotype B (simultaneous mutations of G5 and G6). The 3D-structure of GDF9 protein in A and B haplotypes was rotated 90˚ and 45˚ than wild haplotype, respectively. The missenses G1/p.Arg87His, G4/p.Glu241Lys and G6/p.Val332Ile variants were benign. However both the missenses of G7/p.Val371Met and G8/p.Ser315Phe were probably damaging. Phylogenetic tree of GDF9 gene revealed that individuals with A and B haplotypes were distinct from wild haplotypes with bootstrapping values of 63 and 76, respectively. In conclusion, GDF9 protein in A and B haplotypes showed a higher performance than wild haplotype due to synergism effects of simultaneous mutations. These types of mutations with effect on turn and helix of GDF9 conservative regions showed physical and functional interaction with TGFβ proteins.
بهرهگیری از ژنهای عمده باروری از قبیل GDF9 و BMP15 به بهبود صفات تولیدمثلی در برنامههای اصلاح نژادی گوسفند کمک شایانی خواهد نمود. به منظور ارزیابی جهشهای نقطهای بر روی عملکرد پروتئین، چندشکلی ژنهای GDF9 و BMP15 بر روی 12 گوسفند مهربان به وسیله توالی یابی DNA و مدلسازی ساختار پروتئین بررسی شد. تعداد شش چندشکلی تک نوکلئوتیدی شناخته شده به عنوان جهشهای FecG (G1-G6) در اگزون شماره 1 و 2 ژن GDF9 یافت شد. از میان چندشکلیهای یافت شده، جهش G1 (GDF9 exon1 g.2118 G>A)، G1 (GDF9 exon1 g.2118 G>A) و G6 (GDF9 exon 2 g.3974 G>A) ، جایگزینی اسیدآمینه را در ساختار پروتئین نشان دادهاند. هیچ چندشکلی در اگزون 1 و 2 ژن BMP15 یافت نشد. بر اساس چندشکلیهای شناسایی شده، افراد به سه دسته هاپلوتایپی از قبیل هاپلوتایپ وحشی (بدون جهش)، هاپلوتایپ A (جهشهای همزمان G1، G2، G3 و G4) و هاپلوتایپ B (جهشهای همزمان G5 و G6) تقسیمبندی شدند. ساختار سه بُعدی پروتئین GDF9 در هاپلوتایپ A و B به ترتیب یک چرخش 90 و 45 درجهای را نسبت به هاپلوتایپ نوع وحشی نشان داد. واریانتهای G1/p.Arg87His، G4/p.Glu241Lysو G6/p.Val332Ile بر روی پروتئین بیتأثیر بودند. در حالیکه دو جهشG7/p.Val371Met وG8/p.Ser315Phe به احتمال زیاد تأثیرات مخرب بر روی پروتئین داشتند. بر اساس آنالیز درخت تکامل ژن GDF9، افراد هاپلوتایپ A و B به ترتیب به ارزش خود راه اندازی 63 و 76 از هاپلوتایپ نوع وحشی فاصله داشتند. در نتیجه، پروتین GDF9 در افراد هاپلوتایپ نوع A و B به دلیل اثرات سینرژیسم جهشهای همزمان عملکرد بالاتری را نسبت به افراد هاپلوتایپ نوع وحشی نشان میدهد. این نوع جهشها با تأثیر بر روی مارپیچ و چرخشهای پروتئین GDF9 در نواحی حفاظتی، منجر به اندرکنش فیزیکی پروتئین GDF9 با پروتئینهای فاکتور تبدیل کننده رشد بتا (TGFβ) میشوند.
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