Sequence characterization and promoter identification of porcine APC10 gene

Mol Biol Rep. 2010 Dec;37(8):3841-9. doi: 10.1007/s11033-010-0040-3. Epub 2010 Mar 16.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5' Flanking Region / genetics
  • Amino Acid Sequence
  • Anaphase-Promoting Complex-Cyclosome
  • Animals
  • Base Sequence
  • Conserved Sequence
  • Evolution, Molecular
  • Gene Expression Regulation
  • Genes, Reporter
  • Genome / genetics
  • Humans
  • Luciferases / metabolism
  • Molecular Sequence Data
  • Phylogeny
  • Promoter Regions, Genetic*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sequence Alignment
  • Sequence Analysis, DNA*
  • Sp1 Transcription Factor / metabolism
  • Sus scrofa / genetics*
  • Two-Hybrid System Techniques
  • Ubiquitin-Protein Ligase Complexes / chemistry
  • Ubiquitin-Protein Ligase Complexes / genetics*
  • Ubiquitin-Protein Ligase Complexes / metabolism

Substances

  • RNA, Messenger
  • Sp1 Transcription Factor
  • Luciferases
  • Ubiquitin-Protein Ligase Complexes
  • Anaphase-Promoting Complex-Cyclosome