The Sp family of transcription factors regulates the human laminin α1 gene in JAR choriocarcinoma cells

Abstract

Laminin-111 (α1β1γ1) is the major component of the embryonic and extra-embryonic basement membrane. The laminin α1 chain shows a restricted and developmentally regulated expression in basement membranes of distinct epithelial tissues while β1 and γ1 chains have a wide tissue distribution. To understand how human laminin α1 chain expression is controlled, we cloned and characterized the 5′-flanking region of the human laminin α1 (LAMA1) gene. Transfection studies using serially deleted promoter constructs and JAR choriocarcinoma cells revealed that the minimal promoter fragment resided in the + 31 to − 206 region, which contains a number of GC- and GT/A-rich motifs for the binding of the Sp family of transcription factors. Electrophoretic mobility shift assays and mutational analyses revealed that Sp1 and Sp3 bound specifically to these elements and are important for the promoter activity. Furthermore, we showed that Krüppel-like factors KLF4 and KLF6 also activate transcription of the human LAMA1 gene. Chromatin immunoprecipitation analysis demonstrated recruitment of these transcription factors to the promoter region. These results indicate that transcription of the human LAMA1 gene is controlled by a combination of the actions of Sp1/Sp3 and Krüppel-like factors, KLF4 and KLF6.