Localization, expression and genomic structure of the gene encoding the human serine protease testisin

Hooper, John D.; Bowen, Natalie; Marshall, Heidi; Cullen, Lara M.; Sood, Raman; Daniels, R. J.; Stuttgen, Melanie A.; Normyle, John F.; Higgs, Douglas R.; Kastner, Daniel L.; Ogbourne, Steven M.; Pera, Martin F.; Jazwinska, Elizabeth C.; Antalis, Toni M.

doi:10.1016/s0167-4781(00)00071-3

Cited by 43 publications

(56 citation statements)

References 41 publications

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“…The amplified region spans the proximal promoter, the 5 0 untranslated region, the transcription initiation site and exon I (Hooper et al, 1999(Hooper et al, , 2000. The primers for the first round of PCR were forward 5 0 -TAGTTTGGGTAGAGATTTGGGGAGATTTTT-3 0 (224F), reverse 5 0 -CTCCTACGACTCTACGAAAAACAAAAAATA-3 0 (T845).…”

Section: Rt -Pcr and Real-time Pcr Analysesmentioning

confidence: 99%

“…The human Testisin gene (TEST1; PRSS21) is located within a cluster of serine protease genes on chromosome 16p13.3 (Inoue et al, 1999;Hooper et al, 2000) and consists of six exons ranging in size from 27 to 354 bp interspersed with five introns ranging in size from 97 to 1985 bp. Unusually for serine protease genes, the human Testisin gene contains a 5…”

mentioning

confidence: 99%

“…0 CpG island and a 5 0 CpG rich region, which encompass the proximal promoter, transcription and translation initiation sites, the 5 0 untranslated region and coding/noncoding sequences spanning exons I -III (Hooper et al, 2000). The Testisin 5 0 proximal promoter lacks a TATA consensus sequence, but contains a putative CCAAT box, three consensusbinding sites for Sp1, two potential AP1 binding sites, one putative NFkB binding site and several elements associated with testisspecific expression (Hooper et al, 2000).…”

mentioning

confidence: 99%

“…The Testisin 5 0 proximal promoter lacks a TATA consensus sequence, but contains a putative CCAAT box, three consensusbinding sites for Sp1, two potential AP1 binding sites, one putative NFkB binding site and several elements associated with testisspecific expression (Hooper et al, 2000). Many of these sites contain CpG dinucleotides located wholly within or bordering their sequence that may be affected by CpG methylation.…”

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Hypermethylation of the 5′ CpG island of the gene encoding the serine protease Testisin promotes its loss in testicular tumorigenesis

et al. 2005

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The Testisin gene (PRSS21) encodes a glycosylphosphatidylinositol (GPI)-linked serine protease that exhibits testis tissue-specific expression. Loss of Testisin has been implicated in testicular tumorigenesis, but its role in testis biology and tumorigenesis is not known. Here we have investigated the role of CpG methylation in Testisin gene inactivation and tested the hypothesis that Testisin may act as a tumour suppressor for testicular tumorigenesis. Using sequence analysis of bisulphite-treated genomic DNA, we find a strong relationship between hypermethylation of a 385 bp 5 0 CpG rich island of the Testisin gene, and silencing of the Testisin gene in a range of human tumour cell lines and in 100% (eight/eight) of testicular germ cell tumours. We show that treatment of Testisinnegative cell lines with demethylating agents and/or a histone deacetylase inhibitor results in reactivation of Testisin gene expression, implicating hypermethylation in Testisin gene silencing. Stable expression of Testisin in the Testisin-negative Tera-2 testicular cancer line suppressed tumorigenicity as revealed by inhibition of both anchorage-dependent cell growth and tumour formation in an SCID mouse model of testicular tumorigenesis. Together, these data show that loss of Testisin is caused, at least in part, by DNA hypermethylation and histone deacetylation, and suggest a tumour suppressor role for Testisin in testicular tumorigenesis.

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Section: Rt -Pcr and Real-time Pcr Analysesmentioning

confidence: 99%

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confidence: 99%

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Hypermethylation of the 5′ CpG island of the gene encoding the serine protease Testisin promotes its loss in testicular tumorigenesis

et al. 2005

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“…This clone is part of a contig localizing to chromosome 16p, the same chromosomal arm containing ␣-tryptase (TPS1), ␤-tryptases (e.g. TPSB1), ␥-tryptase (TPSG1), and testisin (PRSS21) genes (7,9,10,18,29). The pancreasin gene appears to reside on the centromeric side of the tryptase locus and on the telomeric side of the testisin locus (i.e.…”

Section: Resultsmentioning

confidence: 99%

Structure and Activity of Human Pancreasin, a Novel Tryptic Serine Peptidase Expressed Primarily by the Pancreas

Bhagwandin

Hau

Clair

et al. 2003

Journal of Biological Chemistry

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In a search for genes encoding the serine peptidases prostasin and testisin, which are expressed mainly in prostate and testis, respectively, we identified a related, novel gene. Sequencing of cDNA allowed us to deduce the full amino acid sequence of the human gene product, which we term "pancreasin" because it is transcribed strongly in the pancreas. The idiosyncratic 6-exon organization of the gene is shared by a small group of tryptic proteases, including prostasin, testisin, and ␥-tryptase. Like the other genes, the pancreasin gene resides on chromosome 16p. Pancreasin cDNA predicts a 290-residue, N-glycosylated, serine peptidase with a typical signal peptide, a 12-residue activation peptide cleaved by tryptic hydrolysis, and a 256-amino acid catalytic domain. Unlike prostasin and other close relatives, human pancreasin and a nearly identical chimpanzee homologue lack a carboxyl-terminal membrane anchor, although this is present in 328-residue mouse pancreasin, the cDNA of which we also cloned and sequenced. In marked contrast to prostasin, which is 43% identical in the catalytic domain, human pancreasin is transcribed strongly in pancreas (and in the pancreatic ductal adenocarcinoma line, HPAC) but weakly or not at all in kidney and prostate. Antibodies raised against pancreasin detect cytoplasmic expression in HPAC cells. Recombinant, epitope-tagged pancreasin expressed in Chinese hamster ovary cells is glycosylated and secreted as an active tryptic peptidase. Pancreasin's preferences for hydrolysis of extended peptide substrates feature a strong preference for P1 Arg and differ from those of trypsin. Pancreasin is inhibited by benzamidine and leupeptin but resists several classic inhibitors of trypsin. Thus, pancreasin is a secreted, tryptic serine protease of the pancreas with novel physical and enzymatic properties. These studies provide a rationale for exploring the natural targets and roles of this enzyme.

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TE2 and TE1 sub‐elements of the testis‐specific histone H1t promoter are functionally different

Wilkerson¹,

Wolfe²,

Grimes³

2003

J of Cellular Biochemistry

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The testis-specific linker histone H1t gene is transcribed exclusively in pachytene primary spermatocytes. Tissue specific expression of the gene is mediated in part by transcriptional factors that bind elements located within the proximal and distal promoter. A 40 bp promoter element, designated H1t/TE, that is located within the proximal promoter between the CCAAT-box and AC-box, is known to be essential for H1t gene transcription in transgenic animals. In the present study, we show by SDS-PAGE analysis of UV crosslinked protein and DNA and by electrophoretic mobility shift assays (EMSA) of testis nuclear proteins separated on a non-denaturing glycerol gradient that the TE1 sub-element is bound by a protein complex. Mutation of TE1 leads to a drop in H1t promoter activity in germinal GC-2spd cells as well as in nongerminal Leydig, NIH3T3, and C127I cell lines. Although TE1 and TE2 sub-elements have similar sequences, mutation of the TE2 sub-element causes an increase in promoter activity in C127I and Leydig cells. The rat TE1 but not TE2 contains a CpG dinucleotide and this cytosine is methylated in liver but not in primary spermatocytes. Methylation of the cytosine at this site almost eliminates nuclear protein binding. Thus, there are significant functional differences in the TE2 and TE1 sub-elements of the H1t promoter with TE1 serving as a transcriptional activator binding site and TE2 serving as a repressor binding site in some cell lines.

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Localization, expression and genomic structure of the gene encoding the human serine protease testisin

Cited by 43 publications

References 41 publications

Hypermethylation of the 5′ CpG island of the gene encoding the serine protease Testisin promotes its loss in testicular tumorigenesis

Hypermethylation of the 5′ CpG island of the gene encoding the serine protease Testisin promotes its loss in testicular tumorigenesis

Structure and Activity of Human Pancreasin, a Novel Tryptic Serine Peptidase Expressed Primarily by the Pancreas

TE2 and TE1 sub‐elements of the testis‐specific histone H1t promoter are functionally different

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