Cell-type-specific interactions at regulatory motifs in the first intron of the lamin A gene

Abstract

Lamins A, C and C2 are alternatively spliced products of the LMNA gene; lamins A and C are expressed in differentiated somatic cells, whereas lamin C2 is expressed in germ cells. We have analyzed a segment of the first intron of the LMNA gene for cell-type-specific regulatory elements. We identified a 420-bp fragment that increased promoter activity in lamin A-expressing cells but repressed activity in undifferentiated cells. DNase I footprinting and electrophoretic mobility shift assays revealed two binding motifs, footprinted region A (FPRA) and FPRB. The hepatocyte nuclear factor-3β was bound to FPRA only in somatic cell extracts and this motif had an inhibitory effect on promoter activity. The retinoic X receptor β, RXRβ, bound near FPRB with extracts from lamin A- or C2-expressing cells, and this site enhanced promoter activity. We have, thus, identified two novel binding sites for transcription factors in a region likely to function as an important regulatory element for the cell-type-specific transcription of A-type lamins.