Cloning of the semenogelin II gene of the rhesus monkey. Duplications of 360 bp extend the coding region in man, rhesus monkey and baboon.

The semenogelin II gene from the rhesus monkey has been cloned and characterized. The transcription unit is split into three exons of 97, 2086 and 124 bp, with two intervening introns of 241 bp and 862 bp. The first exon codes for a 23-amino-acid signal peptide and the two amino-terminal residues of the secreted protein. The second exon codes for the rest of the mature protein, and the third exon contains non-coding nucleotides only. Secreted rhesus monkey semenogelin II consists of 683 amino acid residues, has a calculated M(r) of 77362, is devoid of Cys and Met, and displays a highly repetitive structure composed of ten 60-amino-acid repeats. Hybridization with genomic DNA showed that the semenogelin II gene of man, rhesus monkey and baboon has evolved through extension of the coding region with 360-bp segments. In contrast, the length of the semenogelin I gene of these species appears to be conserved. The two genes are also present in some New World monkeys, as was revealed by hybridization with genomic DNA from the marmoset. However, another New World monkey, the cotton-top tamarin, carries only one semenogelin gene, but also has a gene that is similar to the mouse semenoclotin gene.

The gene of the protease inhibitor SKALP/elafin is a member of the REST gene family.

Members of the REST gene family characteristically have a transcription unit consisting of three exons. The first and the last exon are conserved among members, while the second exon--encoding almost all of the mature protein--differs considerably. The so far known REST genes are highly, and almost exclusively, expressed in the seminal vesicles. By sequence analysis we have now identified the gene for the protease inhibitor SKALP/elafin as a new member of the REST gene family. The protein is expressed in the epidermis and serves, like the product of several REST genes, as substrate for transglutaminase. We have also found what seems to be a locus encompassing both transglutaminases and REST genes centered around the region q12 on the human chromosome 20, raising the question whether the enzymes and substrates have evolved in parallel.

Gene structure of semenogelin I and II. The predominant proteins in human semen are encoded by two homologous genes on chromosome 20.

The genes for semenogelin I and II, the major protein constituents of the human seminal fluid, have been characterized by three overlapping clones in bacteriophage lambda, encompassing 31.5 kilobases (kb) of genomic DNA. The two genes are located 11.5 kb apart in the region q12-q13.1 on chromosome 20. Both genes are relatively compact, spanning only 2.7 and 3.1 kb, respectively. The transcription units are composed of three exons, of which the first encodes the signal peptide, the second encodes the secreted protein, while the third solely contains 3'-noncoding nucleotides. The nucleotide sequences exhibit a similarity of close to 90% in the exons and exceeding 80% in the introns and flanking nucleotides.

Assignment of the human gene for beta-microseminoprotein (MSMB) to chromosome 10 and demonstration of related genes in other vertebrates.

The gene for beta-microseminoprotein MSMB has been studied by DNA hybridization and molecular cloning techniques. Comparative analysis of restriction endonuclease digests of the cloned gene and of leukocyte DNA strongly suggested that the gene is present in a single copy in the haploid human genome. By Southern blot analysis of DNA from somatic cell hybrids, the gene was assigned to chromosome 10. The coding nucleotides of the human gene are separated into four exons by relatively large introns. A related gene might be present in other mammals, birds, and amphibians as revealed by DNA hybridization under conditions of low stringency.

Structure and expression of the human cystatin C gene.

The structural organization of the gene for the human cysteine-proteinase inhibitor cystatin C was studied. Restriction-endonuclease digests of human genomic DNA hybridized with human cystatin C cDNA and genomic probes produced patterns consistent with a single cystatin C gene and, also, the presence of six closely related sequences in the human genome. A 30 kb restriction map covering the genomic region of the cystatin C gene was constructed. The positions of three polymorphic restriction sites, found at examination of digests of genomic DNA from 79 subjects, were localized in the flanking regions of the gene. The gene was cloned and the nucleotide sequence of a 7.3 kb genomic segment was determined, containing the three exons of the cystatin C structural gene as well as 1.0 kb of 5'-flanking and 2.0 kb of 3'-flanking sequences. Northern-blot experiments revealed that the cystatin C gene is expressed in every human tissue examined, including kidney, liver, pancreas, intestine, stomach, antrum, lung and placenta. The highest cystatin C expression was seen in seminal vesicles. The apparently non-tissue-specific expression of this cysteine-proteinase inhibitor gene is discussed with respect to the structure of its 5'-flanking region, which shares several features with those of housekeeping genes.

Molecular cloning of a small prostate protein, known as beta-microsemenoprotein, PSP94 or beta-inhibin, and demonstration of transcripts in non-genital tissues.

In order to study the gene expression of the seminal plasma protein beta-microseminoprotein, also known as PSP94 and beta-inhibin, clones encoding this protein were isolated from a cDNA library constructed in lambda gt11. Nucleotide sequencing confirmed the structure of a previously cloned cDNA. By northern blot analysis identical sized transcripts were demonstrated in the prostate, the respiratory (tracheal, bronchial and lung) tissues and the antrum part of the gastric mucosa. Thus, the protein is not primarily associated with male reproductive function. Although probably of no physiological significance, a slight structural similarity to the ovarian inhibin beta-chains was identified in the C-terminal half of the molecule.