Construction and characterization of a plasmid containing complementary DNA to mRNA encoding the <i>N</i>-terminal amino acid sequence of the rat glutathione transferase Ya subunit

Taylor, John B.; Craig, Roger K.; Beale, D.; Ketterer, Brian

doi:10.1042/bj2190223

Cited by 43 publications

(28 citation statements)

References 32 publications

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“…The rat liver glutathione S-transferase Ya-Yc gene family is comprised of at least five to seven genes (16 (11,(26)(27)(28)(29)(30), the nucleotide differences that exist between the clones are not due to either cloning artifacts or DNA sequencing mistakes. (32).…”

Section: Discussionmentioning

confidence: 99%

Structural analysis of a rat liver glutathione S-transferase Ya gene.

Telakowski-Hopkins¹,

Rothkopf²,

Pickett³

1986

Proc. Natl. Acad. Sci. U.S.A.

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We have isolated and characterized a complete structural gene encoding a rat liver glutathione Stransferase (glutathione transferase; EC 2.5.1.18) Ya subunit. The gene spans =11 kilobases and is comprised of seven exons separated by six introns. A sequence similar to the Goldberg-'Hogness promoter ("TATA" box), TATTA, is located 32 base pairs upstream from the transcription initiation site. Exons 2 and 4 of the glutathione S-transferase gene encode amino acid sequences of the Ya subunit that are highly conserved in the Yc subunit, whereas exons 3 and 5 encode amino acids that are divergent in the Yc subunit. These data suggest that exons 2 and 4 may encode domains of the Ya subunits that have similar structural or functional properties to the corresponding domains in the Yc subunit (e.g., glutathione binding site), whereas exons 3 and 5 may encode domains of the Ya subunit that have unique structural or functional properties to the corresponding domains in the Yc subunit (e.g., substrate binding site).The glutathione S-transferases (glutathione transferase; EC 2.5.1.18) are a family of enzymes that catalyze the conjugation of glutathione to a variety of electrophilic ligands. In addition, the transferases bind heme and bilirubin as well as various exogenous hydrophobic compounds with high affinity (1-3). The enzymes are comprised of binary combinations of at least seven major subunits, Ya, Ya, Ybl, Yb2, Yc, Yn, and Yp, which are electrophoretically distinguishable on one-dimensional NaDodSO4/polyacrylamide gels (4-7). Peptide mapping experiments and carboxyl-terminal sequence analysis have indicated that Ya and Yc subunits are comprised of a mixture of at least two microheterogeneous polypeptides (8-10).Our laboratory has reported the construction of Ya, Yc, Ybl, and Yb2 cDNA clones (11)(12)(13)(14). We have used these clones in RNA blot hybridization and nuclear run-off assays to demonstrate that the rat liver glutathione S-transferase genes are transcriptionally activated by phenobarbital and 3-methylcholanthrene (11,14,15). DNA sequence analysis of the four cDNA clones indicates that the Ya and Yc genes are members of the same gene family, whereas the Yb1 and Yb2 genes are members of a second glutathione S-transferase gene family. Genomic blots using the Ya and Yc cDNAs as probes suggest the presence of at least five to seven Ya and/or Yc genes in the rat (16).In the present study, we have isolated a complete structural gene encoding a Ya subunit of the rat liver glutathione S-transferases. The Ya structural gene spans ""11 kilobases (kb) and is comprised of seven exons separated by six introns. Nucleotide sequence analysis of the exons of the glutathione S-transferase structural gene indicates that the sequence is equivalent to the Ya cDNA clone, pGTB38, which has been isolated and characterized by our laboratory (11). (23). MATERIALS AND METHODS RESULTSIsolation of a Rat Glutathione S-Transferase Structural Gene. In a previous study from our laboratory, we reported Abbreviations: bp, base pair(s); k...

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Section: Discussionmentioning

confidence: 99%

Structural analysis of a rat liver glutathione S-transferase Ya gene.

Telakowski-Hopkins¹,

Rothkopf²,

Pickett³

1986

Proc. Natl. Acad. Sci. U.S.A.

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“…The separated RNA was transferred to Hybond-N membranes (Amersham) by blotting overnight in 20 x SSC. The 32P-labelled antiluteolysin oligonucleotide was hybridized to the RNA blot by a method modified from Taylor, Craig, Beale & Ketterer (1984) by the exclusion of formal¬ dehyde from the hybridization buffer and the use of 5% polyethylene glycol instead of dextran sulphate. Hybridization and washing (to a stringency of 2 x SSC and 0-1% SDS) was carried out at 37°C.…”

Section: Northern Blotmentioning

confidence: 99%

Sheep antiluteolytic interferon: cDNA sequence and analysis of mRNA levels

Stewart

McCann²,

Northrop³

et al. 1989

Journal of Molecular Endocrinology

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A cloned cDNA has been isolated by probing a sheep blastocyst cDNA library using a synthetic oligonucleotide representing the N-terminal amino acid sequence of the antiluteolytic protein, ovine trophoblast protein-1. Sequence analysis of the cDNA confirms the 70% homology between the antiluteolysin and the interferon-alpha family of proteins; however, the sequence reported here differs at several points from previously reported amino acid and cDNA sequences for the antiluteolysin. In-vitro translation of day-16 poly(A)+ RNA indicated that antiluteolysin mRNA is a major constituent of total mRNA at this stage of blastocyst development, and Northern blotting confirmed that antiluteolysin mRNA production occurred between days 13 and 22 after oestrus. This is consistent with the stage at which embryonic extracts are antiluteolytic on administration in vivo. These and other data confirm that the ovine trophoblast antiluteolysin is an interferon, and suggest that at least five isoforms of this protein may exist.

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“…RNA blotting and S1 mapping Analysis of poly(A)-containing RNA by RNA blotting was carried out as described by Taylor et al (1984). Probes were labelled with [a32P]dCVP by nick translation (Rigby et al, 1977).…”

Section: Methodsmentioning

confidence: 99%

In situ hybridisation and S1 mapping show that the presence of infiltrating plasma cells is associated with poor prognosis in breast cancer

Parkes

Collis²,

Baildam³

et al. 1988

Br J Cancer

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Summary In order to identify potential markers of prognosis in breast cancer, representative cDNA libraries were constructed using RNA isolated from primary breast tumour tissue associated with good and poor prognosis. Cross-screening of these libraries repeatedly identified cloned mRNA species associated with the immune system, in particular B-cells, in libraries derived from tumours of poor prognosis. We have used one of these, a KIV light chain cDNA probe, in two complementary studies to investigate the relationship between immunoglobin gene expression and prognosis. The results obtained using a combination of S, mapping, RNA blotting and in situ hybridisation demonstrate that the presence of plasma cells, as defined by infiltrating cells which express high levels of immunoglobulin K-chain mRNA, is associated with a poor prognosis.

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Construction and characterization of a plasmid containing complementary DNA to mRNA encoding the N-terminal amino acid sequence of the rat glutathione transferase Ya subunit

Cited by 43 publications

References 32 publications

Structural analysis of a rat liver glutathione S-transferase Ya gene.

Structural analysis of a rat liver glutathione S-transferase Ya gene.

Sheep antiluteolytic interferon: cDNA sequence and analysis of mRNA levels

In situ hybridisation and S1 mapping show that the presence of infiltrating plasma cells is associated with poor prognosis in breast cancer

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