Nucleotide sequence of two mouse histone H4 genes

Biochemical demonstration of complex formation of histone pre-mRNA with U7 small nuclear ribonucleoprotein and hairpin binding factors.

“…This suggests that the complex forms only very transiently during the processing reaction; this is in agreement with earlier findings that histone RNA processing in vitro begins very rapidly without any detectable time lag (Gick et al, 1986 (Meier et al, 1989) and defined mutants thereof, were synthesized with appropriate 5' and 3' overhanging ends and clones in various combinations between the Sacl and HindII sites of pSP65 (see Figure 2). The relevant nucleotide sequences of the resulting plasmids were determined by the dideoxy method (Sanger…”

“…For this analysis, we used a short pre-mRNA from the H4-12 gene (Meier et al, 1989) containing 49 nt preceding and 36 nt following the processing site, respectively (12/12; Figure 2 histone genes (data not shown). Of eight histone genes analysed, H4-12 is the most efficiently processed in vitro (Streit, 1990).…”

Modifications of protein-DNA interactions in the proximal promoter of a cell-growth-regulated histone gene during onset and progression of osteoblast differentiation.

“…lA. It can be seen that there is extensive sequence similarity between the site II promoter elements of the F0108 human H4 histone gene and the analogous regions of the rat H4 histone genes as well as with other mammalian histone genes, immediately upstream of the TATA box (20,24,30,32,(34)(35)(36). Most importantly, the nucleotides that make specific DNA-protein contacts in vivo are conserved in these sequences (32).…”

Specificities of Caenorhabditis elegans and human hairpin binding proteins for the first nucleotide in the histone mRNA hairpin loop

“…The RNA-binding domain of C. elegans HBP is sufficient to discriminate against binding to mammalian hairpin RNA To know if the difference between human and C. elegans RBD sequences was reflected by different specificities for histone hairpin RNAs, we produced human and C. elegans HBP in vitro and tested these proteins in electrophoretic mobility shift assays (EMSA) with 32 Plabeled RNAs containing either a mammalian or a C. elegans histone mRNA hairpin+ The mammalian hairpin illustrated in Figure 2 as mmHPs RNA is derived from the mouse histone H4+12 gene (Meier et al+, 1989)+ The C. elegans sequence referred to here as ceHP RNA is from the C. elegans H4-3+2 gene (Roberts et al+, 1989)+ The ceHPs RNA, a shorter variant of ceHP RNA, shows the same binding properties as the longer C. elegans substrate but has a reduced propensity to dimerize (Fig+ 2)+ Interestingly, although human HBP bound equally well to the mmHPs and ceHPs RNAs (Fig+ 3B, lanes 2 and 5), the C. elegans HBP bound almost exclusively to ceHPs RNA (Fig+ 3B, lanes 3 and 6)+ This difference remained when HBP fragments lacking either the C-or N-terminal sequences flanking the RBD were used (Fig+ 3A,B)+ Truncated proteins derived from human HBP formed complexes with both mmHPs (Fig+ 3B, lanes 8 and 9) and ceHPs RNA (Fig+ 3B, lanes 13 and 14), whereas complexes with protein fragments derived from C. elegans HBP were only observed with ceHPs RNA (Fig+ 3B, lanes 15 and 16)+ We observed multiple bands with proteins containing the human N-terminus (Fig+ 3B, lanes 2, 5, 8, and 13)+ Some of these complexes may have arisen either from modified FIGURE 1. Sequence alignment of the minimal RNA-binding domains (RBDs) of histone hairpin binding proteins (HBPs)+ The numbering of residues indicated at the top reflects the 82 amino acid RBD peptides of human and C. elegans HBP used in this study+ Amino acids identical in at least three of the five HBPs analyzed are indicated with white text in black boxes and shown again in the consensus sequence+ The minimal 73-amino-acid RBD defined for human HBP by Wang et al+ (1996) is emphasized by a line below the consensus sequence+ Three a-helical regions predicted by the PHDsec program (Rost & Sander, 1993see Martin et al+, 2000) are indicated by double arrows+ ce: C. elegans; hs: human; mm: mouse; xl: X. laevis+ Note that X. laevis has two HBP-like proteins-SLBP1, related to the mammalian nuclear RNA processing factor HBP, and SLBP2, involved in storing translationally inactive maternal histone mRNAs in the oocyte (Wang et al+, 1999)+ FIGURE 2.…”

“…Sequence alignment of the minimal RNA-binding domains (RBDs) of histone hairpin binding proteins (HBPs)+ The numbering of residues indicated at the top reflects the 82 amino acid RBD peptides of human and C. elegans HBP used in this study+ Amino acids identical in at least three of the five HBPs analyzed are indicated with white text in black boxes and shown again in the consensus sequence+ The minimal 73-amino-acid RBD defined for human HBP by Wang et al+ (1996) is emphasized by a line below the consensus sequence+ Three a-helical regions predicted by the PHDsec program (Rost & Sander, 1993see Martin et al+, 2000) are indicated by double arrows+ ce: C. elegans; hs: human; mm: mouse; xl: X. laevis+ Note that X. laevis has two HBP-like proteins-SLBP1, related to the mammalian nuclear RNA processing factor HBP, and SLBP2, involved in storing translationally inactive maternal histone mRNAs in the oocyte (Wang et al+, 1999)+ FIGURE 2. Histone hairpin RNAs used in this work+ The mammalian (mmHPs) and the C. elegans (ceHPs or ceHP) RNA sequences are derived from the mouse H4+12 gene (Meier et al+, 1989) and the C. elegans H4-3+2 gene (Roberts et al+, 1989), respectively+ The ceHPs has a shorter flanking sequence than the ceHP RNA and gray dashed lines indicate the deletion+ The sequence differences between the ceHP and mmHPs RNAs are shown in gray in the C. elegans sequence+ Mutant RNAs: The mm-ceHP has the sequences flanking the mammalian hairpin structure and the C. elegans hairpin sequence; ce-mmHP has the C. elegans flanking sequence and the mammalian hairpin sequence+ ceHP-Uloop and ceHP-CUUCloop RNAs have only one mutation at the fourth and first positions of the C. elegans hairpin sequence, respectively+ FIGURE 3. RNA-binding domains of human and C. elegans HBP bind with similar specificity to the respective full-length proteins to mammalian and C. elegans hairpins+ A: Schematic representation of the hairpin-binding protein+ The central black region is the RBD used in this study+ Double arrows indicate truncated HBP variants lacking either the C-or the N-terminal sequences flanking the RBD+ B: In vitro-binding assays+ The indicated full-length or truncated HBPs made in vitro were incubated with 32 P-labeled mmHPs or ceHPs RNAs as described in Materials and Methods+ The protein-RNA complexes were separated from free RNA by nondenaturing gel electrophoresis (EMSA)+ Free RNAs are shown in lanes 1, 7, and 17 for mmHPs and in lanes 4, 12, and 20 for ceHPs+ Additional complexes obtained with the hs HBP (lanes 2 and 5) and hs Nt-RBD (lanes 8 and 13) preparations are most likely the result of modified proteins or minor translation products+ The mobility difference between the complexes formed by hs RBD with mmHPs (lane 18) or ceHPs RNA (lane 21) may be due to the different sequences and conformations of the two RNAs and was also observed in other experiments (see Figs+ 5 and 6)+ proteins (i+e+, posttranslational modifications) or from minor translation products (i+e+, internal translation initiation), but all of them showed identical binding behavior+ We thus concluded that the structural elements responsible for the discrimination of the C. elegans HBP against binding to mmHPs RNA must be located within its RBD+ This was confirmed in binding studies with the human HBP fragment encompassing the minimal RBD and the corresponding C. elegans protein fragment illustrated in Figure 1+ In this article, we refer to these shorter proteins as human …”