The mouse Brachyury gene and mesoderm formation.
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Animals↗
Biomedical subjects
K Willison
Publications and source records attributed to K Willison.
Mouse preproacrosin: cDNA sequence, primary structure and postmeiotic expression in spermatogenesis.
The primary structure of mouse preproacrosin was deduced by nucleotide sequencing of cDNA clones isolated from a mouse testis cDNA library. The largest cDNA, with 1373 bp, consists of a 11-bp 5'untranslated sequence, a 1254-bp open reading frame terminated by a TGA triplet and a 105-bp 3' untranslated end, including one potential polyadenylation signal. The NH2-terminus of the polypeptide contains a hydrophobic 15-amino acid signal peptide. This cleavable signal sequence is followed by 403 amino acids, representing the acrosin light and the heavy chain of 23 and 380 amino acid residues, respectively. The proteolytic active site segments His, Asp and Ser are part of the heavy chain, as well as a proline-rich COOH-terminus, which is not present in any other serine proteinase studied so far. Furthermore the postmeiotic expression of the preproacrosin gene during mouse spermatogenesis was studied.
Acrosin↗
The t complex polypeptide 1 (TCP-1) is associated with the cytoplasmic aspect of Golgi membranes.
The t complex polypeptide 1 (TCP-1) is a protein of unknown function expressed in large amounts during spermatogenesis. Rat monoclonal antibodies recognizing TCP-1 have been prepared and used to immunoprecipitate and Western blot a 57 kd protein from germ cell and tissue culture cell extracts. In tissue culture cells, indirect immunofluorescent localization of antigen indicated a perinuclear distribution similar to that of the Golgi apparatus. Analysis of the TCP-1 distribution in tissue culture cells showed that the polypeptide was associated with the cytoplasmic aspect of membranes of the trans-Golgi network (TGN). The distribution in spermatids suggested that TCP-1 was localized to structures often associated with the developing acrosome. The TCP-1 antigenic epitopes are highly conserved, allowing the protein to be identified in cells across a wide variety of vertebrate species and tissues. These experiments suggest that TCP-1 may be essential for transport of proteins through the exocytic pathway in all cells and required in large amounts for acrosome formation in developing spermatids.
Acrosome↗
The human homologue of the mouse t-complex gene, TCP1, is located on chromosome 6 but is not near the HLA region.
Southern blot analysis indicates that there are four sequences in the human genome related to the mouse t-complex gene Tcp-1. All four genes were cloned and partial sequencing showed that one of them was a functional gene, and the other three were pseudogenes. The human sequences were all approximately 90% related to each other and 82-89% related to the mouse Tcp-1a sequence. Human TCP1 cDNA clones from both fibrosarcoma and B cell lines confirmed that there was a single expressed gene. mRNA transcripts of different sizes were accounted for by two different polyadenylation signals. The human TCP1 gene shared some amino acid substitutions with the mouse t-complex allele (Tcp-1a) which were not found in Tcp-1b. The functional human TCP1 gene was mapped, using a panel of somatic cell hybrids, as well as in situ analysis, to the long arm of chromosome 6 at 6q23-qter and thus is not closely linked to the HLA complex on the short arm. For this reason and others it is unlikely that there is a human equivalent of the mouse t-complex.
Amino Acid Sequence↗
Molecular analysis of mouse spermatogenesis: isolation of the t-complex polypeptide-1 gene and related sequences.
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Animals↗
The activation of cellular genes in transformed cells.
We have used differential cDNA cloning techniques to isolate a number of genes that are activated as a result of transformation by the DNA tumour virus Simian virus 40. From the nucleotide sequences of the cDNA clones we have been able to identify some of these genes. One of them derives from the major histocompatibility complex and contains a repetitive element that identifies a large number of RNAs present in pluripotential embryonic cells.
Animals↗
Transcripts regulated during normal embryonic development and oncogenic transformation share a repetitive element.
We have previously isolated cDNA clones homologous to mRNAs present at elevated levels in transformed mouse fibroblasts. Clones of Set 1 contain a dispersed repetitive element present thousands of times in the mouse genome. This repeat identifies in mouse embryos a large number of transcripts that are quantitatively regulated during development. At maximal expression these RNAs constitute between 1% and 3% of polyadenylated RNA. A pattern of Set 1-related transcripts very similar to that observed in midgestation embryos is found in pluripotential EC and EK cell lines, and the abundance of these RNAs decreases upon differentiation in vitro. However, the F9 line of EC cells, which has a more restricted developmental capacity, exhibits a much simpler pattern of transcripts containing the Set 1 repeat.
Age Factors↗
The effect of prolonged decompaction on the development of the preimplantation mouse embryo.
A rabbit antiserum to a mouse embryonal carcinoma cell line blocks compaction of cleaving mouse embryos. Cell division is not affected up to the 32-cell stage but intracellular junctions fail to develop. Removal of the antibody at this stage permits compaction to occur and a normal blastocyst develops. Prolonged decompaction beyond the 32-cell embryo results in an increasing proportion of malformed blastocysts in which trophectodermal cells predominate and functional inner cell mass (ICM) cells are reduced or absent. The relationship of compaction to the generation of ICM and trophectoderm lineages in the intact embryo is discussed.
Animals↗
Activation of a Qa/Tla class I major histocompatibility antigen gene is a general feature of oncogenesis in the mouse.
A cDNA clone corresponding to a mRNA present at elevated levels in transformed fibroblasts encodes a Qa/Tla class I major histocompatibility complex (MHC) antigen. High levels of this mRNA are found in all tumour cells tested; the transcript can undergo alternative splicing; and a repetitive sequence within the transcription unit has the characteristics of a transposable element. The immunological implications of MHC gene activation in tumour cells are discussed.
Animals↗
The class I major histocompatibility antigen gene activated in a line of SV40-transformed mouse cells is H-2Dd, not Qa/Tla.
We have previously described several complementary DNA clones isolated because they correspond to messenger RNAs present at higher levels in the simian virus 40 (SV40)-transformed BALB/c 3T3 cell line SV3T3 Cl38 than in the normal, parental BALB/c 3T3 line. One of these clones, pAG64, hybridizes to RNAs which, while present in BALB/c 3T3 cells, are 10-20-fold more abundant in SV3T3 Cl38 and are found at high levels in a wide variety of transformed cell lines. Nucleotide sequence analysis showed that pAG64 encodes a class I antigen of the major histocompatibility complex. To ascertain the identity of pAG64, we compared its sequence with the available sequences of d haplotype class I antigen genes [K locus, L locus, D locus and the Qa gene defined by genomic clone 27.1] and found that it showed multiple clustered differences from each of these sequences. We therefore concluded that it was not derived from the H-2Kd, H-2Ld or H-2Dd genes and thus must correspond to one of the other class I antigen genes, namely those of the Qa/Tla complex, although it was clearly not the Qa gene defined by the genomic clone 27.1. We now report subsequent findings which indicate that pAG64 in fact corresponds to the H-2Dd gene and not to a Qa/Tla gene.
Animals↗
Systematic implementation of an advance health care directive in the community.
In Canada, advance directives have been developed to ensure individual's decisions about health care are known in the event of mental incapacity. This randomized control trial examined the proportion of chronically ill elders receiving Victorian Order of Nurses (VON) services in the home who would complete an advance directive, factors associated with directive completion, treatment choices, and satisfaction with care. The participants consisted of 163 elders with a chronic illness residing within the Hamilton-Wentworth and Haldimand-Norfolk regions in South Central Ontario. Seventy percent of the experimental group completed the directive. Younger patients (p = 0.01) and patients with particular nurses (p = 0.04) were more likely to complete a directive. Psychosocial variables such as mood, depression, and uncertainty in illness did not influence directive completion. Satisfaction with involvement in health care decisions was not changed by this intervention (p = 0.576).
Activities of Daily Living↗