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Cell type‐specific expression of the human RAP1A and RAP1B genes
Author(s) -
Cruise Jennifer,
Niemela Amy,
Augustin Ryan,
Painter Mark,
Hindt Maria
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.728.14
Subject(s) - promoter , biology , gene , reporter gene , microbiology and biotechnology , cell type , cell culture , gene expression , cell , genetics
Despite their important roles in cell adhesion, motility, immune function, and other processes, little is known about the expression control of the Ras‐family small GTPases RAP1A and 1B, located on human chromosomes 1 and 12, respectively. Although altered expression has been noted in a number of cancers and other disease states, no characterization of these genes' basal promoters, or of their proximal or distal regulatory elements has been reported. We examined native expression of both genes in a set of eight human cell lines, using qRT‐PCR. Human umbilical vein endothelial cells (HUVEC) strongly expressed RAP1A, and Saos‐2 osteosarcoma cells were high expressers of RAP1B. MCF‐10A cells expressed both genes at a moderately high level. Promoter‐less GFP reporter constructs and qRT‐PCR were used to identify functional core promoters for each gene. RAP1A constructs identified a sequence upstream of the TSS containing a negative regulatory element that is functional in more than one cell type. While driving GFP expression in other cell types, RAP1B proximal promoter constructs did not function well in Saos‐2 cells, suggesting that the high native expression observed in that line is under the control of distal elements. Three putative distal regulators have been cloned and are under examination.