The complete amino acid sequence of the basic nuclear protein of bull spermatozoa

Coelingh, J.P.; Monfoort, C.H.; Rozijn, Th.H.; Leuven, J.A. Gevers; Schiphof, R.; Steyn-Parvé, Elizabeth P.; Braunitzer, Gerhard; Schrank, B; Ruhfus, A

doi:10.1016/0005-2795(72)90174-2

Cited by 122 publications

(46 citation statements)

References 32 publications

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“…79:177a, 1978), rats (20,21), bulls (10,32,36), boars (36,43), rams (29,36,41), stallions (36), guinea pigs (8,9), rabbits (8,9), and humans (2,12,25,33,39). Complete sequence analysis of the P1 protamines from bulls (10,32), boars (43), rams (41), and humans (2,33) and the predicted amino acid sequence derived from a cDNA sequence for mouse protamine 1, (mPl) (24) have revealed identical lengths of 50 amino acids and strong sequence homologies among these mammalian protamines. P1 protamine is organized into three domains consisting of a central basic core with clusters of arginine and two less basic amino-and carboxyl-terminal regions.…”

mentioning

confidence: 99%

Mouse protamine 2 is synthesized as a precursor whereas mouse protamine 1 is not.

Yelick¹,

Balhorn²,

Johnson³

et al. 1987

Mol. Cell. Biol.

147

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The nuclei of mouse spermatozoa contain two protamine variants, mouse protamine 1 (mPl) and mouse protamine 2 (mP2). The amino acid sequence predicted from mPl cDNAs demonstrates that mPl is a 50-amino-acid protein with strong homology to other mammalian P1 protamines. Nucleotide sequence analysis of independently isolated, overlapping cDNA clones indicated that mP2 is initially synthesized as a precursor protein which is subsequently processed into the spermatozoan form of mP2. The existence of the mP2 precursor was confirmed by amino acid composition and sequence analysis of the largest of a set of four basic proteins isolated from late-step spermatids whose synthesis is coincident with that of mPl. The sequence of the first 10 amino acids of this protein, mP2 precursor 1, exactly matches that predicted from the nucleotide sequence of cDNA and genomic mP2 clones. The amino acid composition of isolated mP2 precursor 1 very closely matches that predicted from the mP2 cDNA nucleotide sequence. Sequence analysis of the amino terminus of isolated mature mP2 identified the final processing point within the mP2 precursor. These studies demonstrated that mP2 is synthesized as a precursor containing 106 amino acids which is processed into the mature, 63-amino-acid form found in spermatozoa.Protamines are small, basic, arginine-rich proteins that replace histones in the later stages of spermatogenesis in many vertebrates (N. B. Hecht, in G. Stein and J. Stein, ed., Basic Chromosomal Proteins: Structure, Organization and Regulation ofthe Gene, in press). During spermatogenesis in mammals, the histones are not directly replaced by protamines but by a population of basic proteins that are transiently associated with the spermatid nucleus (14,35; Hecht, in press). These testis-specific proteins are removed from the condensing chromatin at later stages of spermiogenesis and are replaced by protamines, the predominant class of proteins found complexed with DNA in the mature spermatozoon. Mammalian protamines contain the amino acid cysteine, which is believed to function in the stabilization and compaction of the sperm nucleus through disulfide bond cross-linking (7).Protamines have been isolated from a large number of vertebrates including fish (3, 28), domestic fowl (38), mice (6, 27, 40; A. R.

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mentioning

confidence: 99%

Mouse protamine 2 is synthesized as a precursor whereas mouse protamine 1 is not.

Yelick¹,

Balhorn²,

Johnson³

et al. 1987

Mol. Cell. Biol.

147

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“…Electrophoretic and chromatographic analyses of these proteins have shown that the sperm of certain animals contain multiple protamine amino acid sequence variants and that the number of these variants differs from species to species. The sperm of bulls (24), rams (55,56), and rats (20,50), for example, contain only one protamine variant, whereas mouse sperm have two (6,9) and the sperm of humans (20,78) and certain fish (3) contain three different protamines.…”

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confidence: 99%

“…Although comparisons of the amino acid sequence of bull protamine (24) and the partial sequences of rat (50), boar (69), ram (69), stallion (69), human (34), and mouse (9) protamine indicate that the homology between various mammalian protamines is extensive, the primary structure of fish and mammalian protamines differs considerably . Mammalian protamines all contain a centrally located polyarginine stretch very similar in size to the entire length of the fish protamine, with additional nonbasic amino acid residues located in the 000H-terminal and N112-terminal "tail" fragments.…”

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confidence: 99%

A model for the structure of chromatin in mammalian sperm.

Balhorn¹

1982

The Journal of Cell Biology

510

269

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DNA in mammalian, and most vertebrate sperm, is packaged by protamines into a highly condensed, biochemically inert form of chromatin. A model is proposed for the structure of this DNA-protamine complex which describes the site and mode of protamine binding to DNA and postulates, for the first time, specific inter- and intraprotamine interactions essential for the organization of this highly specialized chromatin. In this model, the central polyarginine segment of protamine binds in the minor groove of DNA, crosslinking and neutralizing the phosphodiester backbone of DNA while the COOH- and NH2-terminal ends of protamine participate in the formation of inter- and intraprotamine hydrogen, hydrophobic, and disulfide bonds. Each protamine segment is of sufficient length to fill one turn of DNA, and adjacent protamines are locked in place around DNA by multiple disulfide bridges. Such an arrangement generates a neutral, insoluble chromatin complex, uses all protamine sulfhydryl groups for cross linking, conserves volume, and effectively renders the chromatin invulnerable to most external influences.

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“…Early studies of mammalian species indicated that there was a shift toward an arginine-rich nucleoprotein during spermiogenesis, and indeed several workers have isolated small basic proteins from spermatozoa. Coelingh et al isolated and sequenced a protein of molecular weight about 6000 from bull spermatozoa which contained large amounts of arginine and cysteine (7,8). Kistler et al have described another nuclear protein (MW about 6200) isolated only from mature rat testes, but which is not present in rat spermatozoa (9,10).…”

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confidence: 99%

New histones found in mature mammalian testes.

Shires¹,

Carpenter²,

Chalkley³

1975

Proc. Natl. Acad. Sci. U.S.A.

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Two new histones have been found in sexually mature testes of several mammals. The new histones have been identified as a new F1 (TFI) and F2b (TF2b) on the basis -of their behavior upon chemical fractionation procedures and electrophoresis at several urea concentrations. The new testis-specific histones are absent in very immature animals and in somatic tissues. However, by day 20, in the rat, the histone pattern is that of the adult, displaying nearly a full complement of TF1 and TF2b. The new histone complement evidently is characteristic of prespermatid germ cells. The degree of evolutionary variations in these histones and in other basic sperm proteins, as detected by change in electrophoretic mobility, appears to be much greater than that seen in somatic histones. The process of spermiogenesis involves the production of a haploid genome, packaged in a highly organized and compact fashion. In echinoderms the compact nucleoprotein contains DNA in association with histones which are either identical or similar to those found in somatic cells (1-4). On the other hand, in arthropods and teleosts somatic histones are replaced in mature spermatozoa by a low molecular weight, arginine-rich class of proteins known as protamines (5, 6). Early studies of mammalian species indicated that there was a shift toward an arginine-rich nucleoprotein during spermiogenesis, and indeed several workers have isolated small basic proteins from spermatozoa. Coelingh et al. isolated and sequenced a protein of molecular weight about 6000 from bull spermatozoa which contained large amounts of arginine and cysteine (7,8). Kistler et al. have described another nuclear protein (MW about 6200) isolated only from mature rat testes, but which is not present in rat spermatozoa (9, 10).We initiated a program designed to compare the basic proteins from spermatozoa of a range of mammals. As Protamines were obtained from the headpieces of mature spermatozoa isolated from the testis, intact epididymis, or semen. The headpieces were separated from the tails of the spermatozoa by blending. During the acid extraction of the histones from the nuclei, the sperm heads were precipitated t To whom reprint requests should be made.in the acid-insoluble fraction. The pelleted sperm heads were washed twice with deionized water, suspended in 15-20 ml (1 ml/g of starting tissue) of 5 M guanidine-hydrochloride, 0.5 M 2-mercaptoethanol, and homogenized. The ensuing gel was adjusted to pH 7 with NaOH and sheared in a Virtis model 45 homogenizer at 70 V for 45 sec. The ionic strength was decreased by adding 2.5 volumes of deionized water. Protamines were extracted in 0.2 M H2SO4 at 40 for 2 hr. After centrifugation at 27,000 X g for 20 min, the supernatant was dialyzed against 0.2 M H2SO4 to remove guanidine-hydrochloride.The bull protamine was obtained from spermatozoa isolated from bull semen, utilizing a slightly modified procedure of Marushige and Marushige (13).Electrophoresis of histones was performed on 15% acrylamide gels as described (14). Elec...

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The complete amino acid sequence of the basic nuclear protein of bull spermatozoa

Cited by 122 publications

References 32 publications

Mouse protamine 2 is synthesized as a precursor whereas mouse protamine 1 is not.

Mouse protamine 2 is synthesized as a precursor whereas mouse protamine 1 is not.

A model for the structure of chromatin in mammalian sperm.

New histones found in mature mammalian testes.

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