Molecular characterisation of theXenopus laevis chaperonin geneCctg

Walkley, Neal A.; Page, Rachel; Malik, Afshan N.

doi:10.1016/s0167-4781(96)88918-4

Cited by 2 publications

(2 citation statements)

References 15 publications

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“…Sequence analysis suggested that these clones encoded at least three genes. The first group of 26 cDNA clones and zebrafish ESTs (EST269466, fb13f04, fc26d03, fc72f03 and fe18d09) from the database of the Washington University zebrafish EST project encoded the γ subunit of zebrafish chaperonin CCT, which shared 87%, 86%, 85%, 70% and 58% amino acid sequence identities with the Xenopus laevis, mouse, human, Drosophila melanogaster and yeast counterparts, respectively (Chen et al, 1994;Kubota et al, 1994;Dunn and Mercola, 1996;Walkley et al, 1996a;Walkley et al, 1996b;Walkley and Malik, 1996). The second group of five cDNA clones encoded a putative protein that had 69% amino acid sequence identity with human C18orf1, a transmembrane protein with a LDL receptor type A domain (Yoshikawa et al, 1998).…”

Section: Transcripts From the Ntn Regionmentioning

confidence: 99%

Identification of chaperonin CCTγ subunit as a determinant of retinotectal development by whole-genome subtraction cloning from zebrafishno tectal neuronmutant

Matsuda

Mishina

2004

Development

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Zebrafish no tectal neuron (ntn) mutant obtained by trimethylpsoralen (TMP) mutagenesis showed defects in tectal neuropil formation and small eyes. We carried out whole-genome subtraction between wild-type and mutant zebrafish embryos using the representational difference analysis (RDA) method. Nineteen subtraction products enabled us to construct genetic and physical maps of the ntn region. Direct selection of cDNAs using a YAC clone encompassing the ntn locus and RT-PCR analysis of transcripts identified a 143 bp deletion in the cct3 gene encoding the γ subunit of chaperonin containing TCP-1 (CCT). Injection of antisense cct3 morpholino oligonucleotides into zebrafish embryos induced characteristic ntn phenotypes including defects in retinal ganglion cell (RGC) differentiation and tectal neuropil formation. Moreover,injection of cct3 mRNA successfully rescued ntn mutant embryos. Our results suggest that RDA is an efficient and widely applicable cloning strategy in zebrafish genetics. The strong expression of the cct3 mRNA started in the entire embryos by 12 hpf and was sustained thereafter, but there were no detectable abnormalities in body patterning and neurogenesis in ntn mutant embryos at 30 hpf. The expression patterns of transcription factor genes ath5 and brn3b that are essential for the development and maintenance of RGCs were indistinguishable between wild-type and ntn mutant embryos, but those of early and late differentiation markers of RGCs, nicotinic acetylcholine receptor β3 and zn5, were diminished in mutant embryos. Immunostaining of acetylated tubulin also revealed the impairment of RGC neurite extension. Thus, the ntnmutation of the cct3 gene impaired the differentiation of retinal neuroepithelial cells to RGCs. Similarly, the expression of brn3b was normal in the tectum of ntn mutants, but tectal neuropil formation was abolished. These results suggest that the γ subunit of chaperonin CCT plays an essential role in retinotectal development.

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Section: Transcripts From the Ntn Regionmentioning

confidence: 99%

Identification of chaperonin CCTγ subunit as a determinant of retinotectal development by whole-genome subtraction cloning from zebrafishno tectal neuronmutant

Matsuda

Mishina

2004

Development

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“…This is a feature that is not shared by the other TpCCT subunits already characterized. Indeed, TpCCTγ and TpCCTη are longer than their counterparts in mammals, Xenopus lae is, Drosophila melanogaster and yeast [6,[33][34][35], with the majority of the extra amino acids (11 residues) being at the C-terminus, a divergent region between CCT subunits. The biological significance of these features of the TpCCT subunits in the function and\or assembly of the Tetrahymena CCT complex remains to be established.…”

Section: Structure Of the Cctα Gene In Tetrahymena And Predicted Amimentioning

confidence: 99%

The third member of the Tetrahymena CCT subunit gene family, TpCCTα, encodes a component of the hetero-oligomeric chaperonin complex

Soares

Cyrne

Casalou

et al. 1997

Biochemical Journal

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The sequence of a third member of the Tetrahymena pyriformis chaperonin CCT ('chaperonin containing TCP1') subunit gene family is presented. This gene, designated TpCCT alpha, is the orthologue of the mouse chaperonin gene TCP1/CCT alpha. To characterize the CCT complex in this ciliate, we have produced polyclonal antibodies against synthetic peptides based on C-terminal sequences deduced from the primary sequences of the TpCCT alpha, TpCCT gamma and TpCCT eta subunits. We have also used polyclonal antibodies produced against recombinant yeast CCT alpha and CCT beta subunits. Using these antibodies, we show that Tetrahymena cells contain a hetero-oligomeric CCT chaperonin comprising at least seven distinct subunits. Three of these were assigned to specific TpCCT genes, whereas a fourth was recognized by the polyclonal antibody against yeast CCT beta, suggesting that this gene is also present in the ciliate. The CCT complex also contains other unidentified proteins that were recognized by the polyclonal antibody UM-1, raised against the putative ATP binding domain of the chaperonin proteins. TpCCT alpha gene expression was shown in exponentially growing cells and cells regenerating their cilia for different periods to have a similar pattern to the previously identified genes TpCCT gamma and TpCCT eta, and also to tubulin genes.

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Molecular characterisation of theXenopus laevis chaperonin geneCctg

Cited by 2 publications

References 15 publications

Identification of chaperonin CCTγ subunit as a determinant of retinotectal development by whole-genome subtraction cloning from zebrafishno tectal neuronmutant

Identification of chaperonin CCTγ subunit as a determinant of retinotectal development by whole-genome subtraction cloning from zebrafishno tectal neuronmutant

The third member of the Tetrahymena CCT subunit gene family, TpCCTα, encodes a component of the hetero-oligomeric chaperonin complex

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