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L M McKay

Publications and source records attributed to L M McKay.

8 recordsLinked to original sources
PubMed1999

Regulation of the Wilms' tumour suppressor protein transcriptional activation domain.

The Wilms' tumour suppressor protein WT1 contains a transcriptional regulatory domain that can either activate or repress transcription depending upon its cellular environment. The mechanistic basis for this dichotomy is unclear however. Here, we dissect the transcriptional regulatory domains of WT1. We find that a region within the domain of WT1 attributed to transcriptional repression is a potent suppressor of the activation domain at several promoters and in different cell types. In vitro transcription analysis suggests that the mechanism of suppression of the activation domain occurs at the level of transcription initiation. Furthermore we find that the WT1 suppression domain is able to inhibit a heterologous activation domain when fused in cis. Dissection of this domain resulted in the delineation of a 30 amino acid region that was sufficient to confer suppression of a transcriptional activation domain both in vivo and in vitro. Additionally, we find that the WT1 transcriptional activation domain interacts with the general transcription factor TFIIB and that this interaction is not affected by the suppression domain. Taken together, these studies suggest that the suppression domain of WT1 interacts with a cosuppressor protein to mediate inhibition of the WT1 transcriptional activation domain.

3T3 Cells↗
PubMed1999

Evolutionary conserved mechanism of transcriptional repression by even-skipped.

Even-skipped (Eve) is a transcriptional repressor involved in segment formation in Drosophila melano-gaster. In order to gain further insights into the mechanism of action of Eve we tested whether it would function as a transcriptional repressor in mammalian cells. We found that Eve was indeed a potent repressor in two different mammalian cell types and at several promoters. In vitro transcription assays confirmed that Eve directly represses transcription initiation when specifically targeted to a promoter. We also found that, unlike the case with transcriptional activators, Eve does not repress transcription synergistically. Analysis of the effect of Eve on preinitiation complex assembly in a crude HeLa cell nuclear extract demonstrated that the Eve repression domain functions by preventing the assembly of TFIID with the promoter. Our data support the hypothesis that Eve contains an active repression domain that functions specifically to prevent preinitiation complex formation.

3T3 Cells↗
PubMed1992

Mapping human X-linked genes in the phalangerid marsupial Trichosurus vulpecula.

We mapped 15 human X-chromosome markers in the common brush-tailed possum, Trichosurus vulpecula (Kerr), which represents the Australian marsupial family Phalangeridae. In situ hybridization was used to localize highly conserved human X-linked genes to chromosomes of T. vulpecula diploid lines. Ten genes located on the long arm of the human X (human Xq genes) all mapped to the possum X chromosome. However, all five genes located on the short arm of the human X (human Xp genes) mapped to autosomes. These findings confirm our previous work, which showed that the X chromosome in macropodid and dasyurid marsupials bears all the human Xq genes but none of the human Xp genes studied. This suggests that the marsupial X is highly conserved, but its gene content reflects that of only part of the eutherian X, a result consistent with our hypothesis that an autosomal region was added to the X early in eutherian divergence.

Animals↗
PubMed1987

Evolution of mammalian X-chromosome inactivation: sex chromatin in monotremes and marsupials.

The inactive mammalian X-chromosome is always late-replicating, and in eutherian mammals it is heterochromatic and hypermethylated. We propose that this multistep system has evolved from a more primitive system, remnants of which may be found in marsupials and monotremes. The heterochromatic X (sex-chromatin body) is a distinctive feature of interphase cells of certain tissues in eutherian females but not males. Thus we have searched for a sex-specific chromatin body in these same tissues in marsupials (brush-tail possum, Trichosurus vulpecula) and monotremes (platypus, Ornithorynchus anatinus), using classical histological techniques. A female-specific chromatin body was observed at low frequency in nuclei of possum corneal epithelium, but not in any other tissues. No sex difference was observed in any monotreme tissue. These data suggest that stabilization of X-chromosome inactivation by heterochromatinization is tissue-specific in marsupials and absent in monotremes.

Animals↗