Sequence characterization and promoter identification of porcine APC10 gene.
APC10 protein, a subunit of the anaphase-promoting complex (APC), plays an essential role in the progression of cells from mitosis to G1. In this study, we cloned and sequenced partial cDNA, intron 1 and 5'-flanking sequences of porcine APC10. The partial cDNA is 595 bp long and has an open reading frame of 558 bp which encodes 185 putative amino acids. Real-time PCR analysis revealed that the porcine APC10 mRNA expression shows a wide distribution and expression levels varies within a small different range in detected tissues. The deduced protein has a high identity with other eight species and comprises a conserved DOC domain. The phylogenetic tree indicated that porcine APC10 has the closest genetic relationship with human, monkey and dog. Promoter activity was demonstrated by transient transfection of 5'-deletion promoter/luciferase constructs into PK15 cells, and indicated that the proximal region from -2,052 to -1,764 is necessary for basal promoter activity. Positive cis-regulatory elements are present from -2,544 to -2,052 and from -3,114 to -2,774, while negative cis-regulatory elements may be present from -2,774 to -2,544. Yeast one-hybrid assay revealed Sp1 can interact with proximal promoter region of porcine APC10.
College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
W B
WB
He
Z W
ZW
Wang
Y
Y
Li
Z L
ZL
Tang
S L
SL
Yang
Y L
YL
Mu
K M
KM
Peng
K
K
Li
D024506:5' Flanking Region / Q000235:genetics; D000595:Amino Acid Sequence; D064173:Anaphase-Promoting Complex-Cyclosome; D000818:Animals; D001483:Base Sequence; D017124:Conserved Sequence; D019143:Evolution, Molecular; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D016678:Genome / Q000235:genetics; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D016329:Sp1 Transcription Factor / Q000378:metabolism; D034421:Sus scrofa / Q000235:genetics*; D020798:Two-Hybrid System Techniques; D043743:Ubiquitin-Protein Ligase Complexes / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D012333:RNA, Messenger; D016329:Sp1 Transcription Factor; D008156:Luciferases; D043743:Ubiquitin-Protein Ligase Complexes; D064173:Anaphase-Promoting Complex-Cyclosome