Genomic organization and molecular characterization of the human ninein gene

Biochem Biophys Res Commun. 2000 Dec 29;279(3):989-95. doi: 10.1006/bbrc.2000.4050.

Abstract

The centrosome plays a key role in the formation of the mitotic spindle, cell polarity, and cell locomotion. Previously we identified a novel centrosomal associated protein hNinein using GSK-3beta as a bait in the yeast two-hybrid assay. In this report, the hNinein genome was found to correspond to 29 exons of genomic sequence on human chromosome 14q22. Promoter analysis predicts that hNinein contains a TATA, two CCAAT, and three GC boxes. The promoter exhibits the following potential transcription factor binding sites: Sp1, p300, and AP-1. In addition, an alternatively spliced isoform, encoded a 2041-amino-acid protein of 237,900 Da, which was designated hNinein-Lm (GenBank AF302773). The hNinein-Lm genome was found to correspond to 28 exons (2'-29). Amino acid sequence comparison with hNinein showed that hNinein-Lm exhibited an EF-hand Ca2+ binding domain in the N-terminus which similar to mouse ninein. Northern blot showed that this hNinein-Lm isoform was expressed more than hNinein in tissues examined. Differential RT-PCR combining Southern blotting also showed that hNinein-Lm is much more abundant compared to hNinein. Two forms of ninein may also imply the status of ninein associated with a pair of the centrioles in the centrosome structure. Furthermore, molecular characterization shows that human ninein is oligomerized at the C-terminal end which overlapped with GSK-3beta binding site, suggesting that oligomerization of ninein may be regulated by GSK-3beta phosphorylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing
  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites
  • Blotting, Northern
  • Blotting, Southern
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism
  • Chromosome Mapping
  • Chromosomes, Human, Pair 14*
  • Cytoskeletal Proteins
  • DNA / analysis
  • Exons
  • GTP-Binding Proteins / genetics*
  • Genome, Human
  • Glycogen Synthase Kinase 3
  • Humans
  • Introns
  • Molecular Sequence Data
  • Nuclear Proteins
  • Protein Structure, Tertiary

Substances

  • Cytoskeletal Proteins
  • NIN protein, human
  • Nuclear Proteins
  • DNA
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Glycogen Synthase Kinase 3
  • GTP-Binding Proteins

Associated data

  • GENBANK/AF302773