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J. Biochem, 2003, Vol. 133, No. 5 615-623
© 2003 Japanese Biochemical Society


CELL

A Novel Rat Orthologue and Homologue for the Drosophila crooked neck Gene in Neural Stem Cells and Their Immediate Descendants

Naoki Amada1,+, Tomoyuki Tezuka1,+,§, Akila Mayeda2, Kazuaki Araki1, Nobuyuki Takei1, Kazuo Todokoro3 and Hiroyuki Nawa1,{ddagger}

1 Molecular Neurobiology, Brain Research Institute, Niigata University, Asahimachi-dori 1-757, Niigata 951-8585; 2 Department of Biochemistry Molecular Biology, University of Miami School of Medicine, Miami, Florida 33136, USA; and 3 Laboratory of Molecular Cell Science, RIKEN institute, Ibaraki, 351-0198

The crooked neck (crn) gene of Drosophila melanogaster encodes a scaffold protein carrying multiple tetratricopeptide repeat (TPR) motifs, and its mutation results in a reduction in the number of neuroblasts and lethality during larval stages. Here, we isolated two structurally related genes from a rat embryonic brain cDNA library. One gene is the rat orthologue of crn, which encodes 690 amino acids including 16 copies of TPR. The other gene, ATH55, encodes an 855 amino acid protein including 21 TPR motifs, which presumably represents a rat crn homologue and an orthologue of human XAB2. Both genes are highly expressed in embryonic brain but their expressions decrease during development. ATH55-like immunoreactivity is present in the ventricular zone and newly formed cortical plate, while CRN-like immunoreactivity is more abundant in a younger ventricular zone. In agreement, both proteins were found to be enriched in cultured neural stem cells and to decrease in response to cell differentiation signals. As indicated for the yeast CRN-like protein, ATH55 and CRN immunoreactivities were both recovered in the nuclear fraction and detected in the splicing complex carrying pre-mRNA. These findings suggest that both TPR-motif-containing proteins are involved in RNA processing of mammalian neural stem cells and their immediate descendants.

+ Both authors contributed equally to this study.

§ Present address: Mitsubishi Chemical Co, Yokohama Res. Ctr., 1000 Kamoshida-cho, Aoba-ku, Yakohama 227.

To whom correspondence should be addressed. E-mail: hnawa{at}bri.niigata-u.ac.jp

{ddagger} Note: Nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number NM139109 for rat ATH55 and NM053797 for rat CRN.


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