Synteny of the genes for thymidine kinase and galactokinase in the mouse and their assignment to mouse chromosome 11

McBreen, P.; Orkwiszewski, Kathryn G.; Chern, Ching J.; Mellman, William J.; Croce, C.M.

doi:10.1159/000130789

Cited by 30 publications

(10 citation statements)

References 14 publications

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“…Cell Culture-143B.TK Ϫ , a human osteosarcoma-derived cell line (ATCC CRL 8303) (12)(13)(14)(15), and hereafter referred to as 143B, was grown in Dulbecco's modified Eagle's medium supplemented with 5 or 10% fetal bovine serum. For induction of apoptosis, semiconfluent dishes of cells were exposed for different times to 1 M staurosporine.…”

Section: Methodsmentioning

confidence: 99%

Mitochondrial Outer Membrane Permeability Change and Hypersensitivity to Digitonin Early in Staurosporine-induced Apoptosis

Duan

Hájek

Lin

et al. 2003

Journal of Biological Chemistry

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We have shown here that the apoptosis inducer staurosporine causes an early decrease in the endogenous respiration rate in intact 143B.TK ؊ cells. On the other hand, the activity of cytochrome c oxidase is unchanged for the first 8 h after staurosporine treatment, as determined by oxygen consumption measurements in intact cells. The decrease in the endogenous respiration rate precedes the release of cytochrome c from mitochondria. Moreover, we have ruled out caspases, permeability transition, and protein kinase C inhibition as being responsible for the decrease in respiration rate. Furthermore, overexpression of the gene for Bcl-2 does not prevent the decrease in respiration rate. The last finding suggests that Bcl-2 acts downstream of the perturbation in respiration. The evidence of normal enzymatic activities of complex I and complex III in staurosporinetreated 143B.TK ؊ osteosarcoma cells indicates that the cause of the respiration decrease is probably an alteration in the permeability of the outer mitochondrial membrane. Presumably, the voltage-dependent anion channel closes, thereby preventing ADP and oxidizable substrates from being taken up into mitochondria. This interpretation was confirmed by another surprising finding, namely that, in staurosporine-treated 143B.TK ؊ cells permeabilized with digitonin at a concentration not affecting the mitochondrial membranes in naive cells, the outer mitochondrial membrane loses its integrity; this leads to a reversal of its impermeability to exogenous substrates. The loss of outer membrane integrity leads also to a massive premature release of cytochrome c from mitochondria. Most significantly, Bcl-2 overexpression prevents the staurosporine-induced hypersensitivity of the outer membrane to digitonin. Our experiments have thus revealed early changes in the outer mitochondrial membrane, which take place long before cytochrome c is released from mitochondria in intact cells.

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Section: Methodsmentioning

confidence: 99%

Mitochondrial Outer Membrane Permeability Change and Hypersensitivity to Digitonin Early in Staurosporine-induced Apoptosis

Duan

Hájek

Lin

et al. 2003

Journal of Biological Chemistry

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“…Two major versions of the enzyme have been characterized in mammalian cells; one is found in the cytoplasm, and the other is found in the mitochondria (8, 27). They are encoded by genes located on different autosomes (15,30,37,63), and although they catalyze the same basic phosphorylation reaction, the biochemical characteristics of each enzyme and of each reaction are unique (27). The cytosolic form of TK is of special interest, since its activity is regulated in association with several cellular metabolic activities and events.…”

mentioning

confidence: 99%

Transcriptional and posttranscriptional mechanisms regulate murine thymidine kinase gene expression in serum-stimulated cells.

Lieberman¹,

Lin²,

Yeh³

et al. 1988

Mol. Cell. Biol.

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We previously isolated and characterized the structure of murine thymidine kinase (tk) genomic and cDNA sequences to begin a study designed to identify regions of the tk gene important for regulated expression during the transition of cells from Go to a proliferating state. In this report, we describe the stable transfection of the cloned gene into L-M(TK-) cells and show that both thymidine kinase (TK) enzyme activity and DNA synthesis increase in parallel when transfectants in Go arrest are stimulated by serum. To define promoter and regulatory regions more precisely, we have constructed a series of tk minigenes and have examined their expression in stable transfectants after serum stimulation. We have identified a 291-base-pair DNA fragment at the 5' end of the tk gene that has promoter function, and we have determined its sequence. In addition, we have found that DNA sequences which mediate serum-induced expression of TK are transcribed, since expression of the murine tk cDNA, fused to a promoter from either the murine tk gene, the simian virus 40 early region, or the herpes simplex virus tk gene, is stimulated by serum. Our constructs also reveal that the murine tk polyadenylation signal is not required for regulation, nor is most of the 3' untranslated region. RNA dot blot analysis indicates that murine cytoplasmic tk mRNA levels always parallel TK enzyme activity. Nuclear runon transcription assays show less than a 2-fold increase in transcription from the cloned tk gene in serumstimulated transfectants, but an 11-fold increase in mouse L929 cells, which are inherently TK+. These results taken together suggest that the murine tk gene is controlled in serum-stimulated cells by a transcriptional mechanism influenced by DNA sequences that flank tk and also by a posttranscriptional system linked to gene sequences that are transcribed.Thymidine kinase (TK; EC 2.7.1.21) participates in the salvage pathway for pyrimidine nucleotide biosynthesis, catalyzing the phosphorylation of thymidine (TdR) to form thymidine 5'-monophosphate. Two major versions of the enzyme have been characterized in mammalian cells; one is found in the cytoplasm, and the other is found in the mitochondria (8, 27). They are encoded by genes located on different autosomes (15,30,37,63), and although they catalyze the same basic phosphorylation reaction, the biochemical characteristics of each enzyme and of each reaction are unique (27). The cytosolic form of TK is of special interest, since its activity is regulated in association with several cellular metabolic activities and events. Enzyme activity is high in cells infected with certain oncogenic viruses (for a review, see reference 29), in many rapidly proliferating tumor cell populations (14), and in cells during passage through the S phase of the cell cycle (3, 35). In contrast, nonproliferating cells, including those that have terminally differentiated (42), and also cells that are not in the S phase (3, 35), express little or no TK activity.We previously described the cloning and phys...

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“…In spite of the greater sensitivity of strain LY-S to the cytotoxic effects of x-radiation and alkylating agents (2)(3)(4)(5), strain LY-S is less mutable than strain LY-R by UV radiation and x-radiation and by alkylating agents at the hypoxanthine (guanine) phosphoribosyltransferase (Hprt) and Na+,K+-ATPase loci (4)(5)(6). In the present work we have compared the mutability of the two strains at the thymidine kinase (Tk) locus situated on chromosome 11 (7,8) using heterozygous Tk+/Tk-strains of LY-R and LY-S. We have found the mutability of the heterozygous LY-S strain (LY-Si) to be as high or higher than that of LY-R heterozygous strains (LY-R16 and LY-R83) at the Tk locus following treatment with x-radiation, UV radiation, or ethyl methanesulfonate (EtMes). Mutant frequencies are much higher at the Tk locus than at the Hprt and Na',K+-ATPase loci for LY-S and LY-R heterozygous strains treated with these agents, with the exception of strain LY-R83, which is poorly mutable at the Tk locus following treatment with x-radiation.…”

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confidence: 99%

Locus specificity in the mutability of L5178Y mouse lymphoma cells: the role of multilocus lesions.

Evans¹,

Mencl²,

Horng³

et al. 1986

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

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Mouse L5178Y strain LY-S and its parental strain LY-R differ in their comparative sensitivities to the cytotoxic effects of various mutagenic agents-i.e., strain LY-S has been found to be more sensitive, less sensitive, or similarly sensitive to individual agents in comparison to strain LY-R. Nevertheless, strain LY-S has been found to be uniformly less mutable than strain LY-R at the hypoxanthine (guanine) phosphoribosyltransferase (Hprt) locus following treatment with x-radiation, UV radiation, or alkylating agents. In the present work we have isolated subclones of strains LY-R and LY-S that are heterozygous at the thymidine kinase (Tk) locus (chromosome 11). We have found that a heterozygous LY-S Tk+/Tk-strain shows as high or higher mutability at the Tk locus than do heterozygous LY-R strains following treatment with x-radiation, UV radiation, or ethyl methanesulfonate.Mutability of all heterozygous strains at the Tk locus is much higher than at the Hprt locus following treatment with these mutagenic agents, with the exception ofone heterozygous LY-R strain that possesses only one chromosome 11 and that is poorly mutable at the Tk locus by x-radiation. On the basis of these results, we have suggested that (i) because of a repair deficiency, multilocus lesions are formed in the DNA of LY-S strains following treatment with radiation or alkylating agents; (ii) multilocus lesions lead to poor recovery of viable mutants when the target locus is closely linked to essential genes and situated on a hemizygous chromosomal region (e.g., the Hprt locus on the X chromosome or the Tk locus in strains monosomic for chromosome 11); and (iii) x-radiation is a relatively poor mutagen at loci situated on hemizygous chromosomal regions, in repair-efficient and repair-deficient cells, because of its tendency to form multilocus lesions.Mouse lymphoma strain L-5178Y-S (LY-S) was first isolated by Alexander and Mikulski (1) following a spontaneous increase in the x-radiation sensitivity ofL5178Y cells growing in vitro. The parental strain was named L5178Y-R (LY-R) to differentiate it from the newly isolated sensitive strain. In spite of the greater sensitivity of strain LY-S to the cytotoxic effects of x-radiation and alkylating agents (2-5), strain LY-S is less mutable than strain LY-R by UV radiation and x-radiation and by alkylating agents at the hypoxanthine (guanine) phosphoribosyltransferase (Hprt) and Na+,K+-ATPase loci (4-6). In the present work we have compared the mutability of the two strains at the thymidine kinase (Tk) locus situated on chromosome 11 (7, 8) using heterozygous Tk+/Tk-strains of LY-R and LY-S. We have found the mutability of the heterozygous LY-S strain (LY-Si) to be as high or higher than that of LY-R heterozygous strains (LY-R16 and LY-R83) at the Tk locus following treatment with x-radiation, UV radiation, or ethyl methanesulfonate (EtMes). Mutant frequencies are much higher at the Tk locus than at the Hprt and Na',K+-ATPase loci for LY-S and LY-R heterozygous strains treated with these age...

show abstract

Synteny of the genes for thymidine kinase and galactokinase in the mouse and their assignment to mouse chromosome 11

Cited by 30 publications

References 14 publications

Mitochondrial Outer Membrane Permeability Change and Hypersensitivity to Digitonin Early in Staurosporine-induced Apoptosis

Mitochondrial Outer Membrane Permeability Change and Hypersensitivity to Digitonin Early in Staurosporine-induced Apoptosis

Transcriptional and posttranscriptional mechanisms regulate murine thymidine kinase gene expression in serum-stimulated cells.

Locus specificity in the mutability of L5178Y mouse lymphoma cells: the role of multilocus lesions.

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