scholarly journals Identification of a G1-S-phase-regulated region in the human thymidine kinase gene promoter.

1990 ◽  
Vol 10 (7) ◽  
pp. 3834-3837 ◽  
Author(s):  
H H Roehl ◽  
S E Conrad
Keyword(s):  
Thymidine Kinase ◽  
Regulatory Region ◽  
Gene Promoter ◽  
S Phase ◽  
Deletion Mutants ◽  
Thymidine Kinase Gene ◽  
Base Pairs ◽  
Kinase Gene ◽  
Transcriptional Start Sites ◽  
Neomycin Resistance

We have identified a regulatory region in the human thymidine kinase gene promoter. A set of promoter deletion mutants was constructed, linked to the bacterial neomycin resistance gene, and stably transfected into Rat3 cells. It was shown that the region between 135 and 67 base pairs upstream of the cap site is required for conveying G1-S-phase regulation to the linked neo gene. In addition, primer extension assays demonstrated that the same transcriptional start sites were used in G1- and S-phase cells and in the various deletion mutants tested.

Identification of a G1-S-phase-regulated region in the human thymidine kinase gene promoter

1990 ◽  
Vol 10 (7) ◽  
pp. 3834-3837
Author(s):  
H H Roehl ◽  
S E Conrad
Keyword(s):  
Thymidine Kinase ◽  
Regulatory Region ◽  
Gene Promoter ◽  
S Phase ◽  
Deletion Mutants ◽  
Thymidine Kinase Gene ◽  
Base Pairs ◽  
Kinase Gene ◽  
Transcriptional Start Sites ◽  
Neomycin Resistance

We have identified a regulatory region in the human thymidine kinase gene promoter. A set of promoter deletion mutants was constructed, linked to the bacterial neomycin resistance gene, and stably transfected into Rat3 cells. It was shown that the region between 135 and 67 base pairs upstream of the cap site is required for conveying G1-S-phase regulation to the linked neo gene. In addition, primer extension assays demonstrated that the same transcriptional start sites were used in G1- and S-phase cells and in the various deletion mutants tested.

scholarly journals G1/S-regulated E2F-containing protein complexes bind to the mouse thymidine kinase gene promoter.

1994 ◽  
Vol 269 (2) ◽  
pp. 1306-1313
Author(s):  
Q.P. Dou ◽  
S. Zhao ◽  
A.H. Levin ◽  
J. Wang ◽  
K. Helin ◽  
...  
Keyword(s):  
Thymidine Kinase ◽  
Protein Complexes ◽  
Gene Promoter ◽  
Thymidine Kinase Gene ◽  
Kinase Gene

scholarly journals Genetic analysis of the human thymidine kinase gene promoter.

1986 ◽  
Vol 6 (8) ◽  
pp. 2903-2909 ◽  
Author(s):  
J A Kreidberg ◽  
T J Kelly
Keyword(s):  
Thymidine Kinase ◽  
Base Pair ◽  
Promoter Region ◽  
Genomic Sequence ◽  
Gene Promoter ◽  
Thymidine Kinase Gene ◽  
Rich Region ◽  
Kinase Gene ◽  
Inverted Orientation ◽  
Pair Sequence

The promoter of the human thymidine kinase gene was defined by DNA sequence and genetic analyses. Mutant plasmids with deletions extending into the promoter region from both the 5' and 3' directions were constructed. The mutants were tested in a gene transfer system for the ability to transform TK- cells to the TK+ phenotype. This analysis delimited the functional promoter to within an 83-base-pair region upstream of the mRNA cap site. This region contains sequences common to other eucaryotic promoters including G X C-rich hexanucleotides, a CAAT box, and an A X T-rich region. The CAAT box is in an inverted orientation and is part of a 9-base-pair sequence repeated twice in the promoter region. Comparison of the genomic sequence with the cDNA sequence defined the first exon of the thymidine kinase gene.

Genetic analysis of the human thymidine kinase gene promoter

1986 ◽  
Vol 6 (8) ◽  
pp. 2903-2909
Author(s):  
J A Kreidberg ◽  
T J Kelly
Keyword(s):  
Thymidine Kinase ◽  
Base Pair ◽  
Promoter Region ◽  
Genomic Sequence ◽  
Gene Promoter ◽  
Thymidine Kinase Gene ◽  
Rich Region ◽  
Kinase Gene ◽  
Inverted Orientation ◽  
Pair Sequence

The promoter of the human thymidine kinase gene was defined by DNA sequence and genetic analyses. Mutant plasmids with deletions extending into the promoter region from both the 5' and 3' directions were constructed. The mutants were tested in a gene transfer system for the ability to transform TK- cells to the TK+ phenotype. This analysis delimited the functional promoter to within an 83-base-pair region upstream of the mRNA cap site. This region contains sequences common to other eucaryotic promoters including G X C-rich hexanucleotides, a CAAT box, and an A X T-rich region. The CAAT box is in an inverted orientation and is part of a 9-base-pair sequence repeated twice in the promoter region. Comparison of the genomic sequence with the cDNA sequence defined the first exon of the thymidine kinase gene.

Ultrastructural Analysis of Bone Marrow Hematopoiesis in Mice Transgenic for the Thymidine Kinase Gene Driven by the IIb Promoter

Blood ◽  
1998 ◽  
Vol 92 (6) ◽  
pp. 2012-2023 ◽  
Author(s):  
Christel Poujol ◽  
Diana Tronik-Le Roux ◽  
Philippe Tropel ◽  
Valérie Roullot ◽  
Alan Nurden ◽  
...  
Keyword(s):  
Thymidine Kinase ◽  
Gene Promoter ◽  
Antigen Expression ◽  
Recovery Phase ◽  
Thymidine Kinase Gene ◽  
Kinase Gene ◽  
Immunogold Staining ◽  
Hematopoietic Stem ◽  
Cd34 Antigen

Transgenic mice have been generated with expression of the herpes virus thymidine kinase gene directed by a 2.7-kb fragment of the IIb murine promoter of the gene encoding the IIb-subunit of the platelet integrin IIbβ3 (Tropel et al, Blood90:2995, 1997). Administration of ganciclovir (GCV) to these mice resulted not only in an acute cessation of platelet production due to the depletion of the megakaryocytic lineage, but also a decrease in erythrocyte and leukocyte numbers. Immunogold staining on ultrathin frozen sections and electron microscopy has now shown that the remaining population of immature hematopoietic cells contain a high proportion of Sca-1+ and CD34+ cells, with CD45R+ cells of the lymphopoietic lineage being maintained. Stromal cells were also preserved. Blood thrombopoietin levels were high. At 4 days of the recovery phase, Sca-1 and CD34 antigen expression decreased with intense proliferation of cells of the three lineages, with megakaryocyte (MK) progenitors being identified by their positivity for glycoprotein IIb-IIIa. These results suggest that transcriptional activity for the IIb gene promoter was present on pluripotent hematopoietic stem cells. At 6 to 8 days after cessation of GCV, numerous mature MK were observed, some of them with deformed shapes crossing the endothelial barrier through thin apertures. Proplatelet production was visualized in the vascular sinus. After 15 days, circulating platelet levels had increased to approximately 65% of normal. Transgenic IIb-tk mice constitute a valuable model to study in vivo megakaryocytopoiesis. © 1998 by The American Society of Hematology.

Ultrastructural Analysis of Bone Marrow Hematopoiesis in Mice Transgenic for the Thymidine Kinase Gene Driven by the IIb Promoter

Blood ◽  
1998 ◽  
Vol 92 (6) ◽  
pp. 2012-2023 ◽  
Author(s):  
Christel Poujol ◽  
Diana Tronik-Le Roux ◽  
Philippe Tropel ◽  
Valérie Roullot ◽  
Alan Nurden ◽  
...  
Keyword(s):  
Thymidine Kinase ◽  
Gene Promoter ◽  
Antigen Expression ◽  
Recovery Phase ◽  
Thymidine Kinase Gene ◽  
Kinase Gene ◽  
Immunogold Staining ◽  
Hematopoietic Stem ◽  
Cd34 Antigen

Abstract Transgenic mice have been generated with expression of the herpes virus thymidine kinase gene directed by a 2.7-kb fragment of the IIb murine promoter of the gene encoding the IIb-subunit of the platelet integrin IIbβ3 (Tropel et al, Blood90:2995, 1997). Administration of ganciclovir (GCV) to these mice resulted not only in an acute cessation of platelet production due to the depletion of the megakaryocytic lineage, but also a decrease in erythrocyte and leukocyte numbers. Immunogold staining on ultrathin frozen sections and electron microscopy has now shown that the remaining population of immature hematopoietic cells contain a high proportion of Sca-1+ and CD34+ cells, with CD45R+ cells of the lymphopoietic lineage being maintained. Stromal cells were also preserved. Blood thrombopoietin levels were high. At 4 days of the recovery phase, Sca-1 and CD34 antigen expression decreased with intense proliferation of cells of the three lineages, with megakaryocyte (MK) progenitors being identified by their positivity for glycoprotein IIb-IIIa. These results suggest that transcriptional activity for the IIb gene promoter was present on pluripotent hematopoietic stem cells. At 6 to 8 days after cessation of GCV, numerous mature MK were observed, some of them with deformed shapes crossing the endothelial barrier through thin apertures. Proplatelet production was visualized in the vascular sinus. After 15 days, circulating platelet levels had increased to approximately 65% of normal. Transgenic IIb-tk mice constitute a valuable model to study in vivo megakaryocytopoiesis. © 1998 by The American Society of Hematology.

Building a metal-responsive promoter with synthetic regulatory elements

1985 ◽  
Vol 5 (6) ◽  
pp. 1480-1489
Author(s):  
P F Searle ◽  
G W Stuart ◽  
R D Palmiter
Keyword(s):  
Thymidine Kinase ◽  
Promoter Region ◽  
Fusion Gene ◽  
Regulatory Elements ◽  
Regulatory Sequence ◽  
Thymidine Kinase Gene ◽  
Coding Region ◽  
Base Pairs ◽  
Promoter Elements ◽  
Kinase Gene

A fusion gene consisting of the promoter region from the mouse metallothionein-I gene joined to the coding region of the herpes simplex virus thymidine kinase gene is efficiently regulated by zinc in a transient assay when transfected into baby hamster kidney cells. Analysis of similar plasmids in which the metallothionein-I promoter region was mutated indicated the presence of multiple metal regulatory elements (MREs) between -176 and -44 base pairs from the cap site. To further investigate the function of MREs, we inserted a synthetic DNA fragment containing the sequence of MRE-a (the element between -55 and -44 base pairs) into the nonresponsive promoter of the thymidine kinase gene in various positions and configurations. Little or no induction by zinc was observed with single insertions of the regulatory sequence, whereas many different constructions having two copies of MRE-a were inducible. The precise position of the two MREs relative to each other or to the thymidine kinase promoter elements had a relatively small effect on the efficiency of induction, but the inducibility could be further increased by the introduction of more MRE-a sequences. MRE-a can function synergistically with the thymidine kinase distal promoter elements, but in the presence of the TATA box alone it functions as a positive, zinc-dependent promoter element.

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