Characterisation of the NUCKS gene on human chromosome 1q32.1 and the presence of a homologous gene in different species

doi:10.1016/j.bbrc.2004.08.153

Biochemical and Biophysical Research Communications

Volume 323, Issue 3, 22 October 2004, Pages 796-801

https://doi.org/10.1016/j.bbrc.2004.08.153 Get rights and content

Abstract

The NUCKS gene is located on human chromosome 1q32.1 and consists of seven exons and six introns. The gene lacks a TATA box but contains two Inr elements, two GC boxes, and one consensus-binding site for E2F-1. NUCKS is expressed in all human adult and foetal tissues investigated, and has all the features of being a housekeeping gene. Both data searches and Western immunoblotting experiments show that a homologous protein is present in fish, amphibians, and birds but not in insects and yeast, suggesting that NUCKS is a vertebrate specific gene. In all the species investigated, the protein contains several consensus phosphorylation sites for cyclin-dependent kinases and CK-2, and we have shown that the fish protein (like mammalian NUCKS) indeed is a substrate for CDK1 and CK-2 in vitro. The NUCKS protein is also conserved with respect to a DNA-binding domain previously characterised in mammals, and two putative bipartite nuclear localisation signals.

Section snippets

Materials and methods

PCR amplification of introns. A BAC clone (RP11-212H11) containing the NUCKS gene was obtained from The Wellcome Sanger Institute. DNA from the BAC clone was isolated by aid of the Large-Construct kit (Qiagen) as described by the manufacturer. The small and medium-sized introns were confirmed by PCR with Platinum Taq DNA Polymerase High Fidelity (Invitrogen) utilising DNA from the BAC clone as template and primers corresponding to sequences in the different exons.

In an attempt to confirm the

Results and discussion

A query of the working draft sequence of the Human Genome Project database with the sequence of the full-length NUCKS cDNA, using the BLAST program at NCBI (http://www.ncbi.nlm.nih.gov/genome/seq/HsBlast.html), yielded two regions on chromosome 1, locus 1q32.1, containing the cDNA sequence. Computer analysis revealed that the human NUCKS gene contains seven exons and six introns (Fig. 1A). Exon I is separated from exon II by a very long intron of approximately 20 kbp, while the rest of the

Acknowledgments

This work was supported by the Norwegian Cancer Society, Anders Jahre’s Foundation for the Promotion of Sciences, the Astri and Birger Torsted Legacy, and the Blix Legacy. CDK1/Cyclin B was a kind gift from L. Meijer, CNRS, Cell Cycle Group, Station Biologique, Roscoff, France, and CK-2 was a kind gift from Lorenzo Pinna, Dipartimento di ChimicaBiologica, University of Padova, Italy. The chicken liver was a gift from Prior, Norway, the Xenopus laevis oocytes were from Department of Molecular

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Characterisation of the NUCKS gene on human chromosome 1q32.1 and the presence of a homologous gene in different species

Abstract

Section snippets

Materials and methods

Results and discussion

Acknowledgments

FEBS Lett.

Biochem. Biophys. Res. Commun.

Arch. Biochem. Biophys.

Biochim. Biophys. Acta

J. Mol. Biol.

J. Biol. Chem.

A novel, highly phosphorylated protein, of the high-mobility group type, present in a variety of proliferating and non-proliferating mammalian cells

Eur. J. Biochem.

The ubiquity of the highly phosphorylated nuclear protein P1. A novel, highly phosphorylated protein, of the high-mobility group type, present in a variety of proliferating and non-proliferating mammalian cells

Biochem. Int.

Phosphorylation of the high-mobility-group-like protein P1 by casein kinase-2

Eur. J. Biochem.