Isolation and characterization of a mouse fully replication-dependent H1 gene within a genomic cluster of core histone genes

• Published in: The Journal of biological chemistry 1987-12, Vol.262 (35), p.17118-17125
• Main Authors: Yang, Y S, Brown, D T, Wellman, S E, Sittman, D B
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Biotechnology; DNA Replication; DNA Restriction Enzymes - metabolism; Fundamental and applied biological sciences. Psychology; Genes; Genetic engineering; Genetic technics; Histones - genetics; Methods. Procedures. Technologies; Mice; Molecular and cellular biology; Molecular cloning; Molecular genetics; Molecular Sequence Data; Nucleic Acid Hybridization; Oligonucleotides; Replication; RNA, Messenger - analysis
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)45499-3

We have used an oligonucleotide complementary to a sequence coding for the conserved central globular domain of H1s to screen a mouse genomic library for H1 genes. We then used a series of universal histone oligonucleotides to identify five different H1 genes which were linked to core histone genes. We characterized one of the H1 genes which was linked to an H2a, an H2b, an H3, and an H4 histone gene. This characterization involved: 1) sequencing of the coding region of the gene and several hundred base pairs of flanking region. 2) Comparison of this sequence to other H1 sequences from other organisms. This sequence analysis clearly showed that the gene coded for an H1 and identified H1 consensus sequences in the 5′- and 3′-flanking region. 3) Mapping of the 5′- and 3′-ends of the mRNA complementary to this gene by S1 nuclease analysis. 4) Identifying this gene and an adjacent H3 gene as being of the fully replication-dependent expression class, by measuring changes in the steady state levels of their mRNAs in the presence of hydroxyurea and during differentiation of murine erythroleukemia cells.