Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.7.1.21 (thymidine kinase)
 7,561 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have isolated a cDNA that encodes the murine CCAAT-binding protein mYB-1. The deduced amino acid sequence shows 95% identity with its presumed human homologue (hYB-1A) which was originally isolated as a protein that binds to the Y box of MHC class II genes. In vitro translated mYB-1 binds to CCAAT boxes of the MHCIIE alpha, HSVTK and mouse PCNA promoters but not to alpha-globin or human thymidine kinase CCAAT boxes. Interestingly, complexes formed between the in vitro translated protein and the various CCAAT boxes display the property of being competed more efficiently with self competitor DNA, regardless of the CCAAT box initially used as a probe. A similar phenomenon was observed in a cell extract of Con-A stimulated murine splenocytes when the same competition assays were performed. These results may reflect the generation of multiple forms of a particular CCAAT-binding protein, such as mYB-1, that display distinct, yet overlapping, DNA binding specificities.
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PMID:Unusual DNA binding characteristics of an in vitro translation product of the CCAAT binding protein mYB-1. 174 Dec 93

NF-Y is a sequence-specific DNA-binding protein that recognizes the Y box, a promoter element common to all major histocompatibility complex class II genes. Since the 14-base Y element harbors a CCAAT box in reverse, we were prompted to ask whether NF-Y is actually a CCAAT box-binding protein and whether it is related to the previously described CCAAT-binding factors CBP and CTF/NF-I. Data from gel retardation, methylation interference, saturation mutagenesis, and cross-competition experiments establish definitively that NF-Y is an entirely distinct CCAAT box-binding entity. Moreover, these experiments have uncovered a fourth CCAAT-binding protein, NF-Y(star) that interacts with the thymidine kinase promoter. Clearly, then, there exists a multiplicity of factors that recognize CCAAT sequences; it now becomes imperative to understand the functional significance of this multiplicity.
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PMID:A multiplicity of CCAAT box-binding proteins. 347 5

The nuclear protein interacting with the distal CCAAT box of human thymidine kinase (TK) gene promoter has been suggested to be a specific TK-CCAAT-binding protein, which is responsible for the serum-dependence of TK transactivation in normal human IMR-90 fibroblasts. By biochemical characterization, TK-CCAAT-binding protein was found to be distinct from other known CCAAT-binding proteins (Pang, J. H., and Chen, K. Y. (1993) J. Biol. Chem. 268, 2909-2916). In this study, we identify NF-Y, which is composed of Ya and Yb subunits, to be responsible for the TK-CCAAT binding activity in the crude nuclear extract from HL-60 cells. The interaction of NF-Y with the distal CCAAT box of the TK promoter in the crude extract appeared to be more heat-sensitive than that in the DNA affinity chromatography purified fraction. We have further established that the serum dependence of TK-CCAAT binding activity in normal IMR-90 fibroblasts is due to the decrement of NF-Ya, but not NF-Yb expression following serum-deprivation, and that such serum dependence is absent in HL-60 cells.
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PMID:Human thymidine kinase CCAAT-binding protein is NF-Y, whose A subunit expression is serum-dependent in human IMR-90 diploid fibroblasts. 802 44

CBP/tk, CCAAT Binding Protein for thymidine kinase, has been shown to bind to the distal and proximal CCAAT elements in human TK gene at G1/S boundary in normal human IMR-90 cells after serum stimulation (Pang and Chen, 1993). We now show that the serum-induced binding activity of CBP/tk was inversely related to the population doubling level (PDL) of the normal IMR-90 cells. However, little or almost no CBP/tk binding activity was observed in cells derived from patients with premature aging syndromes (e.g., Werner, Hutchinson-Gilford, and Cockayne syndrome). In contrast, CBP/tk binding activity in SV-40 virus-transformed human cells and in HeLa cells was overexpressed at levels 5- to 15-fold higher than that in normal cells and appeared to be deregulated. The half-life of CBP/tk binding activity in SV-40 transformed cells was at least 10 times longer than that in normal IMR-90 cells, suggesting that posttranslational control may contribute to the deregulation. CBP/tk binding activity detected in other mammalian cells such as murine NIH3T3, an immortal cell line, did not reveal any cell cycle dependence either. Further characterization of CBP/tk binding complex indicates that the binding complex may contain NF-YA and NF-YB and that the binding activity was sensitive to oxidizing reagents. Taken together, our data showed that the age- and cell cycle-dependent nature of CBP/tk is a function of cell types and that CBP/tk binding activity may be subjected to posttranslational and redox regulation.
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PMID:The age-dependent binding of CBP/tk, a CCAAT binding protein, is deregulated in transformed and immortalized mammalian cells but absent in premature aging cells. 870 9

Expression of thymidine kinase gene in normal human diploid cells is both cell cycle- and age-dependent and appears to be transcriptionally regulated. Strong DNA protein binding with a 28-bp fragment (-91/-64) that contains the distal inverted CCAAT box is observed in serum-stimulated young (low population doubling level) IMR-90 cells but not in senescent cells. This cell cycle- and age-dependent binding factor was termed CBP/ tk, indicating CCAAT binding protein for the thymidine kinase gene. Based on immunoshift assay and purification, it has been suggested that CBP/tk is equivalent to NF-Y, previously identified as the binding protein for the Y box within E alpha gene promoter. In this study, we examined the mRNA level and protein amount of NF-Y, proteins in young and old IMR-90 cells during serum stimulation by Northern and Western blot blot analyses. In addition, we compared (1) the turnover rate of NF-Y in IMR-90 cells with that of CBP/tk binding activity and (2) the levels of NF-Y and CBP/tk in normal and cancer cells. Both NF-YA and NF-YB were constitutively expressed at mRNA level in IMR-90 cells. However, expression of NF-YA, and to a lesser degree, NF-YB, at the protein level were clearly age-dependent. The half-life of NF-YA and NF-YB were, respectively, 4- and > 10-fold longer than that of CBP/tk binding activity in IMR-90 cells. In addition, we found that the amount of NF-Y did not correlate with the overexpression of CBP/tk binding activity in HeLa cells. Taken together, our results suggested that although CBP/tk may contain NF-YA or related proteins, NF-A and NF-YB alone may not account for all the characteristics of CBP/tk observed in normal and transformed human cells.
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PMID:Cell cycle- and age-dependent transcriptional regulation of human thymidine kinase gene: the role of NF-Y in the CBP/tk binding complex. 886 61