Expression of the mouse WNK1 gene in correlation with ganglioside GD3 and functional analysis of the mouse WNK1 promoter.
WNK1 is one of WNK (With No K=Lysine) protein kinases which comprise a newly described subfamily. Our studies showed that expression of the mouse WNK1 gene was dramatically suppressed in a tumor cell line when its phenotype was altered by suppression of the GD3-synthase gene expression. The mouse WNK1 gene was expressed at a high level at early stage of embryonic brain and its expression decreased as brain developed, similar to the expression pattern of the GD3-synthase gene. To study transcriptional regulation, we cloned a 5'-flanking 1239-bp fragment of the mouse WNK1 gene. This fragment contains a number of potential consensus binding sites for transcription factors, including Sp1, AP2, CCAAT, Est-1, Oct-1, CNBP, and NFkB, but lacks a TATA box. Primer extension identified multiple putative transcriptional initiation sites, including several sites downstream of the ATG codon. Activities of the promoter fragments were assessed in mouse breast Sa/R-MT cells by transient transfection and the results showed that the promoter elements between -700 and -977 is required for maintaining a high level of promoter activity of the TATA-less mouse WNK1 gene.
Zeng
Guichao
G
Institute of Molecular Medicine and Genetics, Medical College of Georgia, 1120 15th Street CB-2803, Augusta, GA 30912, USA. gzeng@mail.mcg.edu
Gao
Luoyi
L
Xia
Tian
T
Gu
Yihua
Y
Yu
Robert K
RK
D024506:5' Flanking Region / Q000235:genetics; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D001921:Brain / Q000196:embryology / Q000378:metabolism; D045744:Cell Line, Tumor; D005732:Gangliosides / Q000378:metabolism*; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D015972:Gene Expression Regulation, Neoplastic; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D015778:Minor Histocompatibility Antigens; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012799:Sialyltransferases / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D024363:Transcription Initiation Site; D014158:Transcription, Genetic; D014162:Transfection; D000076204:WNK Lysine-Deficient Protein Kinase 1
D005732:Gangliosides; D015778:Minor Histocompatibility Antigens; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; C026226:ganglioside, GD3; D008156:Luciferases; D012799:Sialyltransferases; C043662:alpha-N-acetylneuraminate alpha-2,8-sialyltransferase; D017346:Protein Serine-Threonine Kinases; D000076204:WNK Lysine-Deficient Protein Kinase 1; C498744:Wnk1 protein, mouse