Molecular Analysis of Alternative Transcripts of the Equine Cordon-Bleu WH2 Repeat Protein-Like 1 (COBLL1) Gene.
The purpose of this study was to investigate the alternative splicing in equine cordon-bleu WH2 repeat protein-like 1 (COBLL1) gene that was identified in horse muscle and blood leukocytes, and to predict functional consequences of alternative splicing by bioinformatics analysis. In a previous study, RNA-seq analysis predicted the presence of alternative spliced isoforms of equine COBLL1, namely COBLL1a as a long form and COBLL1b as a short form. In this study, we validated two isoforms of COBLL1 transcripts in horse tissues by the real-time polymerase chain reaction, and cloned them for Sanger sequencing. The sequencing results showed that the alternative splicing occurs at exon 9. Prediction of protein structure of these isoforms revealed three putative phosphorylation sites at the amino acid sequences encoded in exon 9, which is deleted in COBLL1b. In expression analysis, it was found that COBLL1b was expressed ubiquitously and equivalently in all the analyzed tissues, whereas COBLL1a showed strong expression in kidney, spinal cord and lung, moderate expression in heart and skeletal muscle, and low expression in thyroid and colon. In muscle, both COBLL1a and COBLL1b expression decreased after exercise. It is assumed that the regulation of COBLL1 expression may be important for regulating glucose level or switching of energy source, possibly through an insulin signaling pathway, in muscle after exercise. Further study is warranted to reveal the functional importance of COBLL1 on athletic performance in race horses.
Department of Animal Science, College of Natural Resources and Life Science, Pusan National University, Miryang 627-706, Korea.
Jeong-Woong
JW
Park
College of Pharmacy, Dankook University, Cheonan 330-714, Korea .
Hyun-Jun
HJ
Jang
Department of Animal Science, College of Natural Resources and Life Science, Pusan National University, Miryang 627-706, Korea ; Life and Industry Convergence Research Institute, College of Natural Resources and Life Science, Pusan National University, Miryang 627-706, Korea .
Sangsu
S
Shin
Department of Animal Science, College of Natural Resources and Life Science, Pusan National University, Miryang 627-706, Korea.
Hyun-Woo
HW
Cho
Department of Animal Science, College of Natural Resources and Life Science, Pusan National University, Miryang 627-706, Korea.
Jae-Young
JY
Choi
Subtropical Animal Experiment Station, National Institute of Animal Science, Rural Development Administration, Jeju 690-150, Korea .
Nam-Young
NY
Kim
Genomic Informatics Center, Hankyong National University, Anseong 456-749, Korea .
Hak-Kyo
HK
Lee
Department of Equine Sciences, Sorabol College, Gyeongju 780-711, Korea .
Kyong-Tak
KT
Do
Genomic Informatics Center, Hankyong National University, Anseong 456-749, Korea .
Ki-Duk
KD
Song
Department of Animal Science, College of Natural Resources and Life Science, Pusan National University, Miryang 627-706, Korea ; Life and Industry Convergence Research Institute, College of Natural Resources and Life Science, Pusan National University, Miryang 627-706, Korea .
Byung-Wook
BW
Cho
Alternative Splicing; Athletic Performance; COBLL1; Horse; Muscle; RNA-seq