Molecular and functional analyses of the fast skeletal myosin light chain2 gene of the Korean oily bitterling, Acheilognathus koreensis.
We identified and characterized the primary structure of the Korean oily bitterling Acheilognathus koreensis fast skeletal myosin light chain 2 (Akmlc2f), gene. Encoded by seven exons spanning 3955 bp, the deduced 168-amino acid AkMLC2f polypeptide contained an EF-hand calcium-binding motif and showed strong homology (80%-98%) with the MLC2 proteins of Ictalurus punctatus and other species, including mammals. Akmlc2f mRNA was highly enriched in skeletal muscles, and was detectable in other tissues. The upstream regions of Akmlc2f included a TATA box, one copy of a putative MEF-2 binding site and several putative C/EBPβ binding sites. The functional activity of the promoter region of Akmlc2f was examined using luciferase and red fluorescent protein reporters. The Akmlc2f promoter-driven reporter expressions were detected and increased by the C/EBPβ transcription factor in HEK293T cells. The activity of the promoter of Akmlc2f was also confirmed in the developing zebrafish embryo. Although the detailed mechanism underlying the expression of Akmlc2f remains unknown, these results suggest the muscle-specific expression of Akmlc2f transcript and the functional activation of Akmlc2f promoter by C/EBPβ.
Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Korea. heejkong@hotmail.com
Hee Jeong
HJ
Kong
Ye-Ji
YJ
Lee
Woo-Jin
WJ
Kim
Hyung Soo
HS
Kim
Bong-Seok
BS
Kim
Cheul Min
CM
An
Sang-Yeob
SY
Yeo
Hyun Kook
HK
Cho
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D024722:Cardiac Myosins / Q000235:genetics*; D002460:Cell Line; D003530:Cyprinidae / Q000235:genetics*; D004247:DNA / Q000032:analysis; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism*; D018994:Myosin Light Chains / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D056910:Republic of Korea; D016415:Sequence Alignment; D016385:TATA Box / Q000235:genetics; D015027:Zebrafish / Q000196:embryology / Q000235:genetics
D022782:CCAAT-Enhancer-Binding Protein-beta; C545012:CEBPB protein, human; D018994:Myosin Light Chains; D012333:RNA, Messenger; C107184:myosin light chain 2; D004247:DNA; D024722:Cardiac Myosins; D002118:Calcium