Yeast two-hybrid screens implicate DISC1 in brain development and function

doi:10.1016/j.bbrc.2003.10.101

Biochemical and Biophysical Research Communications

Volume 311, Issue 4, 28 November 2003, Pages 1019-1025

https://doi.org/10.1016/j.bbrc.2003.10.101 Get rights and content

Abstract

DISC1 is a candidate gene for involvement in the aetiology of major psychiatric illnesses including schizophrenia. We report here the results of DISC1 yeast two-hybrid screens using human foetal and adult brain libraries. Twenty-one proteins from a variety of subcellular locations were identified, consistent with observations that DISC1 occupies multiple subcellular compartments. The cellular roles of the proteins identified implicate DISC1 in several aspects of central nervous system development and function, including gene transcription, mitochondrial function, modulation of the actin cytoskeleton, neuronal migration, glutamate transmission, and signal transduction. Intriguingly, mutations in one of the proteins identified, WKL1, have been previously suggested to underlie the aetiology of catatonic schizophrenia.

Section snippets

Methods

Vector construction. A full-length human DISC1 open reading frame corresponding to sequence with GenBank Accession No. AF222982 was amplified by nested PCR from human foetal heart cDNA using Advantage-GC cDNA polymerase mix (Clontech) and inserted into pDBLeu. This construct is referred to as pDBLeuDISC1. The cloned DISC1 open reading frame was fully sequenced to confirm sequence integrity.

Yeast two-hybrid library screen. Library-scale competent MaV203 yeast cells (Gibco-BRL) were

Results and discussion

Table 1, Table 2 list the proteins that were identified in this yeast two-hybrid screen. To exclude incorrect identification of interactors resulting from the presence of more than one prey plasmid in the same yeast colony, all proteins listed were confirmed by plasmid extraction, sequencing, and subsequent retransformation back into yeast, followed by assay of pair-wise interactions.

When full-length DISC1 was used as bait, 19 different proteins were identified from the adult brain library

Summary

We have identified potential DISC1-interacting proteins that implicate DISC1 in a range of functions including gene transcription, mitochondrial function, modulation of the actin cytoskeleton, neuronal migration, glutamate transmission, and signal transduction. Overall, it is becoming apparent that although DISC1 lacks any known functional motifs, it is likely to be critical to development and functioning of the brain by acting as a scaffold to bring together certain proteins, such as signal

Acknowledgements

We thank Pippa Thomson for critical reading of this manuscript. This work was supported by the Caledonian Research Foundation, the Stanley Medical Research Institute, and the Scottish Hospitals Endowment Research Trust.

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Citation Excerpt :
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Yeast two-hybrid screens implicate DISC1 in brain development and function

Abstract

Section snippets

Methods

Results and discussion

Summary

Acknowledgements

Lancet

Am. J. Hum. Genet.

Genomics

Genomics

Exp. Cell Res.

J. Biol. Chem.

J. Biol. Chem.

Neuron

Neuron

Neuron

Biochem. Biophys. Res. Commun.

FEBS. Lett.

J. Biol. Chem.

Mol. Cell. Neurosci.

FEBS Lett.

Neuron

Neuron

Neuron

J. Biol. Chem.

Curr. Opin. Neurobiol.

Arch. Biochem. Biophys.

J. Biol. Chem.

The neurodevelopmental hypothesis of schizophrenia: following a trail of evidence from cradle to grave

Dev. Psychopathol.