Limits...
The promoter for intestinal cell kinase is head-to-head with F-Box 9 and contains functional sites for TCF7L2 and FOXA factors.

Sturgill TW, Stoddard PB, Cohn SM, Mayo MW - Mol. Cancer (2010)

Bottom Line: The ICK promoter contains consensus motifs for several FOX-family transcription factors that align when mouse and human are compared using EMBOSS.A minimal promoter for ICK contains functional sites for beta-catenin/TCF7L2 and FOXA.These data are consistent with functions that have been proposed for ICK in development and in proliferation or survival of some breast and colon cancers.

View Article: PubMed Central - HTML - PubMed

Affiliation: Departments of Pharmacology and Internal Medicine, University of Virginia, 1300 Jefferson Park Avenue, Charlottesville, Virginia 22908, USA. tws7w@virginia.edu

ABSTRACT

Background: Intestinal cell kinase (ICK; GeneID 22858) is a conserved MAPK and CDK-like kinase that is widely expressed in human tissues. Data from the Cancer Genome Anatomy Project indicated ICK mRNA is increased in cancer, and that its expression correlated with expression of mRNA for an uncharacterized F-box protein, FBX9 (GeneID: 26268). ICK and FBX9 genes are arranged head-to-head on opposite strands, with start sites for transcription separated by approximately 3.3 kb. We hypothesized ICK and FBX9 are potentially important genes in cancer controlled by a bidirectional promoter.

Results: We assessed promoter activity of the intergenic region in both orientations in cancer cell lines derived from breast (AU565, SKBR3), colon (HCT-15, KM12), and stomach (AGS) cancers, as well as in embryonic human kidney (HEK293T) cells. The intergenic segment was active in both orientations in all of these lines, and ICK promoter activity was greater than FBX9 promoter activity. Results from deletions and truncations defined a minimal promoter for ICK, and revealed that repressors and enhancers differentially regulate ICK versus FBX9 promoter activity. The ICK promoter contains consensus motifs for several FOX-family transcription factors that align when mouse and human are compared using EMBOSS. FOXA1 and FOXA2 increase luciferase activity of a minimal promoter 10-20 fold in HEK293T cells. Consensus sites for TCF7L2 (TCF4) (Gene Id: 6934) are also present in both mouse and human. The expression of beta-catenin increased activity of the minimal promoter approximately 10 fold. ICK reference mRNAs (NM_014920.3, NM_016513) are expressed in low copy number and increased in some breast cancers, using a ten base tag 5'-TCAACCTTAT-3' specific for both ICK transcripts.

Conclusion: ICK and FBX9 are divergently transcribed from a bidirectional promoter that is GC-rich and contains a CpG island. A minimal promoter for ICK contains functional sites for beta-catenin/TCF7L2 and FOXA. These data are consistent with functions that have been proposed for ICK in development and in proliferation or survival of some breast and colon cancers.

Show MeSH

Related in: MedlinePlus

FBX9 and ICK are divergently transcribed from a bidirectional promoter. Equal numbers of SKBR3 and AU565 cells were seeded into 96-well plates, transfected with the indicated constructs (Figure 1), then assayed for luciferase activity in each well by the methods described. Data in figures 2 and 3 were obtained by the same procedures. Bar, ± S.D.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2876993&req=5

Figure 2: FBX9 and ICK are divergently transcribed from a bidirectional promoter. Equal numbers of SKBR3 and AU565 cells were seeded into 96-well plates, transfected with the indicated constructs (Figure 1), then assayed for luciferase activity in each well by the methods described. Data in figures 2 and 3 were obtained by the same procedures. Bar, ± S.D.

Mentions: The full intergenic segment (constructs ICK-1 and FBX9-1) was active in both orientations in all six of the lines, suggesting that ICK and FBX9 share a bidirectional promoter. Analyses in the different lines (Figs. 2, 3, 4) show elements in the common SspIb to PstIb fragment are important for bidirectional activity, and may account for the correlated expression of FBX9 and ICK in microarray data that motivated this study. Our analyses (Figs. 2, 3, &4) show that the intergenic segment is not a constitutive, bidirectional promoter because the FBX9 activity relative to ICK activity is variable.


The promoter for intestinal cell kinase is head-to-head with F-Box 9 and contains functional sites for TCF7L2 and FOXA factors.

Sturgill TW, Stoddard PB, Cohn SM, Mayo MW - Mol. Cancer (2010)

FBX9 and ICK are divergently transcribed from a bidirectional promoter. Equal numbers of SKBR3 and AU565 cells were seeded into 96-well plates, transfected with the indicated constructs (Figure 1), then assayed for luciferase activity in each well by the methods described. Data in figures 2 and 3 were obtained by the same procedures. Bar, ± S.D.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2876993&req=5

Figure 2: FBX9 and ICK are divergently transcribed from a bidirectional promoter. Equal numbers of SKBR3 and AU565 cells were seeded into 96-well plates, transfected with the indicated constructs (Figure 1), then assayed for luciferase activity in each well by the methods described. Data in figures 2 and 3 were obtained by the same procedures. Bar, ± S.D.
Mentions: The full intergenic segment (constructs ICK-1 and FBX9-1) was active in both orientations in all six of the lines, suggesting that ICK and FBX9 share a bidirectional promoter. Analyses in the different lines (Figs. 2, 3, 4) show elements in the common SspIb to PstIb fragment are important for bidirectional activity, and may account for the correlated expression of FBX9 and ICK in microarray data that motivated this study. Our analyses (Figs. 2, 3, &4) show that the intergenic segment is not a constitutive, bidirectional promoter because the FBX9 activity relative to ICK activity is variable.

Bottom Line: The ICK promoter contains consensus motifs for several FOX-family transcription factors that align when mouse and human are compared using EMBOSS.A minimal promoter for ICK contains functional sites for beta-catenin/TCF7L2 and FOXA.These data are consistent with functions that have been proposed for ICK in development and in proliferation or survival of some breast and colon cancers.

View Article: PubMed Central - HTML - PubMed

Affiliation: Departments of Pharmacology and Internal Medicine, University of Virginia, 1300 Jefferson Park Avenue, Charlottesville, Virginia 22908, USA. tws7w@virginia.edu

ABSTRACT

Background: Intestinal cell kinase (ICK; GeneID 22858) is a conserved MAPK and CDK-like kinase that is widely expressed in human tissues. Data from the Cancer Genome Anatomy Project indicated ICK mRNA is increased in cancer, and that its expression correlated with expression of mRNA for an uncharacterized F-box protein, FBX9 (GeneID: 26268). ICK and FBX9 genes are arranged head-to-head on opposite strands, with start sites for transcription separated by approximately 3.3 kb. We hypothesized ICK and FBX9 are potentially important genes in cancer controlled by a bidirectional promoter.

Results: We assessed promoter activity of the intergenic region in both orientations in cancer cell lines derived from breast (AU565, SKBR3), colon (HCT-15, KM12), and stomach (AGS) cancers, as well as in embryonic human kidney (HEK293T) cells. The intergenic segment was active in both orientations in all of these lines, and ICK promoter activity was greater than FBX9 promoter activity. Results from deletions and truncations defined a minimal promoter for ICK, and revealed that repressors and enhancers differentially regulate ICK versus FBX9 promoter activity. The ICK promoter contains consensus motifs for several FOX-family transcription factors that align when mouse and human are compared using EMBOSS. FOXA1 and FOXA2 increase luciferase activity of a minimal promoter 10-20 fold in HEK293T cells. Consensus sites for TCF7L2 (TCF4) (Gene Id: 6934) are also present in both mouse and human. The expression of beta-catenin increased activity of the minimal promoter approximately 10 fold. ICK reference mRNAs (NM_014920.3, NM_016513) are expressed in low copy number and increased in some breast cancers, using a ten base tag 5'-TCAACCTTAT-3' specific for both ICK transcripts.

Conclusion: ICK and FBX9 are divergently transcribed from a bidirectional promoter that is GC-rich and contains a CpG island. A minimal promoter for ICK contains functional sites for beta-catenin/TCF7L2 and FOXA. These data are consistent with functions that have been proposed for ICK in development and in proliferation or survival of some breast and colon cancers.

Show MeSH
Related in: MedlinePlus