Protamine messenger RNA: Evidence for early synthesis and accumulation during spermatogenesis in rainbow trout

Iatrou, Kostas; Spira, Arthur W.; Dixon, Gordon H.

doi:10.1016/0012-1606(78)90062-3

Cited by 64 publications

(29 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…4B). In trout, synthesis of the protamine mRNA is known to occur during the primary spermatocyte stage (22). The testis-specific 3-kb SOX-LZ mRNA increased in quantity as the protamine mRNA.…”

Section: Resultsmentioning

confidence: 99%

A Gene That Is Related to SRY and Is Expressed in the Testes Encodes a Leucine Zipper-Containing Protein

Takamatsu

Kanda

Tsuchiya

et al. 1995

Molecular and Cellular Biology

View full text Add to dashboard Cite

SRY-related cDNA encoding a protein with a high-mobility-group (HMG) box and a leucine zipper motif, which was designated SOX-LZ, was isolated from a rainbow trout testis cDNA library. Comparison of this cDNA with the mouse homologous cDNA isolated from a testis cDNA library exhibits an overall amino acid sequence identity of 77%, which is in striking contrast to the abrupt loss of amino acid sequence homology outside the HMG box found among mammalian SRY genes. In both rainbow trout and mice, Northern (RNA) blot analyses have revealed the presence of a testis-specific 3-kb-long SOX-LZ mRNA, and this transcript appeared coincidentally with the protamine mRNA, suggesting its expression in the germ line. A recombinant HMG box region protein encoded by SOX-LZ could bind strongly with an oligonucleotide containing an AACAAT sequence, which is also recognized by mouse Sry and Sox-5. Upon cotransfection into CHO cells, In fish, sex determination and differentiation are attributed to both genetic and environmental factors (4,7,45). With regard to salmonids, male heterogamety has been demonstrated cytologically for several species (30,39,40), and a Y chromosome-specific DNA fragment has been isolated (11), although their sex chromosomes show little morphological differentiation, suggesting an early stage of heteromorphic evolution. Genetic evidence has also indicated that an XY system operates to control sex determination (6,21,24). However, in contrast to mammals, sex determination does not appear to be so strictly bound to the sex chromosomes, and functional sex inversion can be induced in both directions by treatment with exogenous sex steroids (12,20). Thus, fish provide a particularly intriguing biological system in which to study sex determination and differentiation. SOX-LZSRY, a candidate gene responsible for testis determination, has been isolated from the sex determining regions of human and mouse Y chromosomes (16,36). Analyses of XY sexreversed females (2,18,19,23) and testis formation of mice transgenic for Sry (27) have equated SRY and the testis-determining factor. SRY encodes a protein with a DNA-binding motif known as the high-mobility-group (HMG) box (16,36). Surprisingly, comparison of mammalian SRY sequences reveals that sequence conservation is largely confined to the HMG box, suggesting its rapid evolution (13,16,36,41,43). In addition to SRY, genes encoding an SRY type HMG box (SOX genes) have been identified by PCR with primers based on the conserved amino acid sequences of the HMG boxes of mammalian SRY (5,9,10,15,29,42,44). To investigate sex determination and differentiation in fish, we have isolated cDNA clones for a rainbow trout SOX gene (SOX-LZ) expressed in testes. From a phylogenetic interest, we have also isolated mouse homologous cDNA clones. SOX-LZ genes of both species show not only an overall structural conservation but also similar mRNA expression patterns. MATERIALS AND METHODSAmplification of rainbow trout SRY-related HMG box sequence. Total RNA was extracted from rainbow trout t...

show abstract

Section: Resultsmentioning

confidence: 99%

A Gene That Is Related to SRY and Is Expressed in the Testes Encodes a Leucine Zipper-Containing Protein

Takamatsu

Kanda

Tsuchiya

et al. 1995

Molecular and Cellular Biology

View full text Add to dashboard Cite

show abstract

“…This variation in total RNA yield was probably due to extreme increases in certain transcripts related to development of germ cells (also see Liu et al 2000, Kusakabe et al 2002a. For example, protamine is a nuclear protein that is synthesized only in the spermatid cells of the testis and substitutes for histones in sperm chromatin during the haploid phase of spermatogenesis (Gilmour & Dixon 1972, Iatrou & Dixon 1978. Protamine transcripts increased 300-400 times in rainbow Figure 6 Changes in mRNA levels of FSH-R, LH-R, StAR, P450scc, P450C17, P45011 , 3 -HSD, 11 -HSD and 20 -HSD during spermatogenesis.…”

Section: Discussionmentioning

confidence: 99%

Changes in mRNAs encoding steroidogenic acute regulatory protein, steroidogenic enzymes and receptors for gonadotropins during spermatogenesis in rainbow trout testes

Kusakabe¹,

Nakamura²,

Evans³

et al. 2006

Journal of Endocrinology

110

View full text Add to dashboard Cite

In vertebrates, sperm development and maturation are directly regulated by gonadal steroid hormone secretion. The relationships among the expression of genes encoding steroidogenic proteins and receptors for gonadotropins, and testicular steroid production have not yet been comprehensively determined in male teleosts. In this study, the changes in levels of mRNAs encoding follicle-stimulating hormone (FSH) receptor, luteinizing hormone (LH) receptor, steroidogenic acute regulatory protein (StAR), cytochrome P450 cholesterol side-chain cleavage, 3 -hydroxysteroid dehydrogenase/n5-4-isomerase, cytochrome P450 17 -hydroxylase/17,20-lyase, cytochrome P450 11 -hydroxylase, 11 -hydroxysteroid dehydrogenase and 20 -hydroxysteroid dehydrogenase were determined by real-time, quantitative PCR assays and related to changes in serum steroid levels throughout the reproductive cycle in male rainbow trout. Serum 11-ketotestosterone and 17 ,20 -dihydroxy-4-pregnen-3-one levels were measured by RIA. Although the pattern of change in the mRNA levels for the enzymes was variable, the increases in steroidogenic enzyme mRNAs started prior to a significant increase of serum steroid levels. The patterns of transcript levels of FSH and LH receptors suggest that changes in StAR and steroidogenic enzyme transcripts are largely mediated by the FSH receptor during early and mid-spermatogenesis and by the LH receptor during late spermatogenesis and spermiation. Levels of StAR (10-fold) and P450 17 -hydroxylase/ 17,20-lyase (sevenfold) transcripts changed with the greatest magnitude and were closely related to the changes in serum steroids, suggesting that changes in StAR and P450 17 -hydroxylase/17,20-lyase abundance are likely to be the major influences on overall steroidogenic output during the reproductive cycle in male rainbow trout.

show abstract

“…3). The low extent of annealing observed at high Cot values (<103) probably represents the reaction of some genes that were transcribed early in spermatogenesis to produce a stable mRNA [i.e., protamine genes (13)] and that were then inactivated by condensation into nucleoprotamine. The DNA-driven cross-reassociation reactions (Figs.…”

Section: Methodsmentioning

confidence: 99%

Limited action of micrococcal nuclease on trout testis nuclei generates two mononucleosome subsets enriched in transcribed DNA sequences.

Levy-Wilson¹,

Dixon²

1979

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

View full text Add to dashboard Cite

Hybridization experiments show that DNA extracted from two distinct subsets of mononucleosomes (MN1 and MN2) generated by a limited action of micrococcal nuclease on trout testis nuclei is enriched t7-fold in sequences that are transcribed into cytoplasmic polyadenylated RNA in trout testis cells. Both subsets of mononucleosomes contain eight core histones, but MN1 also possesses one or two molecules of a small, basic, high-mobility-group (HMG) protein H6 [Levy W., B., Connor, W. & Dixon, G. H. (1979) J. BioL Chem. 254, 609-620], bound to a DNA fragment of 140 base pairs. In contrast, MN2 contains 1 molecule of HI but no H6, and its DNA length is somewhat longer at 140-190 base pairs. The preferential release of these two subsets of mononucleosomes is correlated with the presence of a second larger HMG protein, HMG-T, in the linker regions flanking both types of mononucleosomes. The HMG-T-containing linker regions appear to be considerably more susceptible to attack by micrococcal nuclease than Hicontaining linkers. Cross-reassociation reactions between the DNA from MN1 and MN2 subsets indicate that they share a significant extent of sequence overlap but also that each subset contains specific sequences that are absent in the other subset.We have recently reported that limited digestion of trout testis nuclei by micrococcal nuclease releases preferentially two kinds of mononucleosomes, MN1 and MN2 (Fig. 1). MN1 monosomes are soluble in 0(.1 M NaCl, lack H1, and contain the four inner histones together with one or two molecules of the smallest trout high-mobility-group (HMG) Preparation of Total Trout DNA and RNA. The procedures for the preparation of total trout DNA and polyadenylated cytoplasmic RNA have been described in detail (2, 5, 6).Preparation of Unlabeled DNA from Various Chromatin Subfractions. DNA was prepared from MN1, MN2, oligosomes, and nucleoprotamine fractions by extraction with phenol/chloroform, followed by chloroform/isoamyl alcohol as described (2).For DNA-driven hybridization reactions, all DNA samples were incubated for 30 min at 37°C with RNase A at 20,g/ml followed by deproteinization with phenol/chloroform and recovery of the DNA fragments from the aqueous phase.In The publication costs of this article were defrayed in part by page charge payment. This particle must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

show abstract

Protamine messenger RNA: Evidence for early synthesis and accumulation during spermatogenesis in rainbow trout

Cited by 64 publications

References 61 publications

A Gene That Is Related to SRY and Is Expressed in the Testes Encodes a Leucine Zipper-Containing Protein

A Gene That Is Related to SRY and Is Expressed in the Testes Encodes a Leucine Zipper-Containing Protein

Changes in mRNAs encoding steroidogenic acute regulatory protein, steroidogenic enzymes and receptors for gonadotropins during spermatogenesis in rainbow trout testes

Limited action of micrococcal nuclease on trout testis nuclei generates two mononucleosome subsets enriched in transcribed DNA sequences.

Contact Info

Product

Resources

About