Cloning and expression of murine hematopoietic specific chromatin remodeling gene SMARCA5

Mouse erythroleukemia cell line was used to generate a library of genes expressed during erythroid differentiation induced by N,N′-hexamethylene-bis-acetamide (HMBA). By subtraction hybridization we detected 63 genes encoding DNA binding molecules, chromatin remodeling genes, signaling proteins, translation regulating factors, metabolic enzymes, mitochondrial genes and expressed sequence tags (EST). The differential gene expression pattern was further confirmed by northern blotting. We used mouse model of accelerated erythropoiesis induced by phenylhydrazine and carbon monoxide-induced hypoxia. The differential expression kinetics of subtracted genes was found during in vivo accelerated erythropoiesis but not during hypoxia. We studied the tissue-specific expression of the ESTs and out of them cloned a murine cDNA of a novel hematopoietic specific chromatin remodeling gene SMARCA5 from cDNA library. Moreover, SMARCA5 is expressed in two transcriptional variants: 4 and 5 kb. In MEL cells and in phenylhydrazine-stimulated mice, SMARCA5 was silenced in hematopoietic cells entering the genetic program of erythroid differentiation. We screened a murine genomic BAC library and isolated SMARCA5 gene which is being further analyzed. Our results demonstrating differentially expressed genes in erythropoiesis are supported by recent studies [Gubin et al. Genomics 59(2):168–77, (1999)] on primary erythroid cells stimulated by erythropoietin.