Functional analysis of the 5' promoter region of the rat lamin A gene

Abstract

The A-type lamins are constituents of the nuclear lamina in differentiated cells and have been proposed to play an important role in nuclear organization. In this study, we isolated and characterized a genomic clone containing the putative promoter region of the rat lamin A gene. Sequence analysis of about 2 kb of this region combined with primer extension data revealed the presence of a TATA box at -33, a GC box at -101, and AP1 motifs at -7, -424, and -1677. Deletion analysis of the promoter fragments in three mammalian cell lines indicated that a 221-bp segment of the proximal promoter containing the GC box and AP1 motif at -7 was sufficient to give high levels of luciferase activity in reporter gene assays. Mutations in these two motifs resulted in considerable loss of reporter gene activity. Analysis by electrophoretic mobility shift assays (EMSAs) has provided evidence for specific binding of the AP1 and Sp1 family of transcription factors to the promoter, a conclusion supported by DNase I footprinting data. This characterization of the 5' promoter region of the lamin A gene should afford a basis for the further clarification of the mechanism of regulation of this important gene during growth and development.