The Primary Structure of Histone H1 from Sperm of the Sea Urchin Parechinus angulosus. 1. Chemical and Enzymatic Fragmentation of the Protein and the Sequence of Amino Acids in the Four N-Terminal Cyanogen Bromide Peptides

Abstract

The primary structure of the amino-terminal 84 residues of sperm histone H1p,,,,,tinus has been determined. The sequence is : Pro -Gly -Ser-Pro-Gln-Lys-Arg -Ala-Ala-Ser-Pro -Arg -Lys-Ser -ProArg -Lys-Ser -Pro -Lys-Lys-Ser-Pro -Arg-Lys -Ala-Ser -Ala-Ser -Pro -Arg -Arg -Lys -Ala-Lys -ArgAla-Arg -Ala-Ser-Thr -Of the five major species of histones found in most eukaryotic organisms, histone H1 is the only one that does not form part of the core particle structure of the chromosome [ l -51. Histones H1 may be involved in the higher-order structure of chromosomes since they are active agents in salt-induced chromatin condensation [6].Histones H5 found in erythrocytes of birds and reptiles have many homologies in their amino acid sequences to histone H1 [7]. Histones H1 and H5 are the most variable among the histones and in many cases H1 histones show species and tissue specificity [8,9]. In general H1 and H5 histones appear to consist of three domains [lo, 111. At the amino-terminal end there is a variable length of random coil. In rabbit thymus H1 [12] and in calf thymus H1 [13] the aminoterminal 40 residues are rich in alanine, lysine and proline. A similar distribution has been found in the first 26 residues of trout testis H1 [14]. In contrast the amino-terminal random-coil regions of avian H5 histones are shorter and not particularly basic except for a short region from residues 12 to 22 [7]. Following the variable-sized amino-terminal random coil there is a hydrophobic region of approximately 80 residues in all H1 and H5 species whose structures have been studied [12-151. This region has a low basic amino acid content, few alanine residues and no proline, and probably exists in vivo as a globular region [lo, 111. At the carboxyl end of all H1 species studied, there are regions of approximately 100 residues with a very high content of lysine, alanine and proline. The same situation occurs in H5 proteins except that, in addition, there are a large number of arginine residues present. A group of lysine, arginine, alanine-rich histones from sperm cells of marine invertebrates [16,17] have been studied. These proteins have been called dsl proteins, but no information on their primary structure is available. The ds1 protein from the sea urchin Arbacia lixula has an almost identical amino acid composition to that of histone H1 from sperm cells of Parechinus angulosus as described in the present paper. Puigdomenech et al. [17] further conclude that the arginine residues in the ds protein are located at both ends of the molecule. Most of the arginine residues of Parechinus HI histone are also located at both ends of the molecule (see also the following paper [18]). In the present paper the purification and isolation of the protein and peptides of the entire protein and the amino acid sequence of the NHz-terminal 84 residues are reported, while the complete primary structure is reported and its unique features are discussed in the following paper [ 181. MATERIALS AND METHODS MaterialsCollection of sperm cells, cold stor...

DNA‐binding sperm proteins with oligo‐arginine clusters function as potent activators for egg CK‐II

“…3B). Polypeptides a, b, d, e and f were identified as histones HI, H2A, H2B s, H3 and H4, respectively, since their partial N-terminal amino acid sequences (a: PGSPQKRAASP-RKSPRKSPKKS; b: SGRGKGAKGKAKAKSRSSRA; d: PRSPAKTSPRKGSPRKGSPRKGSPSRKASP; e: ARTKQ-TARKSTGGKAPRKQL; and f: SGRGKGGKGLGKGGA-KRHRK) corresponded exactly to the sequences of these five histones from gonads and spermatozoa of Parechinus angulosus [16,17] and Psammechinus miliaris [18]. In addition, the Nterminal 28 residue sequence (PRSPSKTSPRKGSPRKGSPS-RKASPKRG) of polypeptide c purified from spermatozoa of H. pulcherrimus was 96.4% homologous to the analogous sequence of histone H2B 2 of another species, Parechinus angulosus: alanine at position 5 was substituted with serine in H. pulcherrimus [19].…”

Mr determination of histones from their electrophoretic mobility in acidic urea gels in the absence of detergents

“…charge per 10000 Da. This figure is in most proteins close to the mol% basic amino acids present ' Ele~trophoretic mobility was measured relative to histone H4 in 15% acrylamide-8 M urea gels, pH 2.3 d Corrected mobility was calculated by dividing the relative mobility by the cationic charge using the charge of histone H4 as a reference given in the table f Error in the M, determination is the difference between the estimated and the actual weight expressed as a percentage g Histone H4 is heterogeneous with respect to the number of E-N-acetylated lysine residues present [7,10] Sperm histones have been isolated from the sea urchin Purechinus angulosus [8]. CN, cyanogen bromide cleavage fragment [S].…”

“…The isolation procedures of the various histones have been summarized in [7]. Sea urchin sperm Hl was isolated and cleaved with CNBr as described by Strickland et al [8] and histone H3 according to Brandt and Von Holt [9].…”

Sequence of mRNA coding for bindin, a species-specific sea urchin sperm protein required for fertilization.

“…The observations reported here confirm that sexual determination in sea urchins does indeed involve differential expression of genes in male and female gonads, and they provide a specific molecular marker useful for further studies ofthis aspect of developmental gene regulation. Other evidence (37)(38)(39) demonstrates that there are also H1 and H2B histone genes that are expressed specifically in testis, the products of which are found only in sperm nuclei. Thus there is evidently a battery of malespecific genes that are activated during the differentiation of the sperm and are otherwise quiescent.…”