Online reference database of European Y-chromosomal short tandem repeat (STR) haplotypes.

The reference database of highly informative Y-chromosomal short tandem repeat (STR) haplotypes (YHRD), available online at http://ystr.charite.de, represents the largest collection of male-specific genetic profiles currently available for European populations. By September 2000, YHRD contained 4688 9-locus (so-called "minimal") haplotypes, 40% of which have been extended further to include two additional loci. Establishment of YHRD has been facilitated by the joint efforts of 31 forensic and anthropological institutions. All contributing laboratories have agreed to standardize their Y-STR haplotyping protocols and to participate in a quality assurance exercise prior to the inclusion of any data. In view of its collaborative character, and in order to put YHRD to its intended use, viz. the support of forensic caseworkers in their routine decision-making process, the database has been made publicly available via the Internet in February 2000. Online searches for complete or partial Y-STR haplotypes from evidentiary or non-probative material can be performed on a non-commercial basis, and yield observed haplotype counts as well as extrapolated population frequency estimates. In addition, the YHRD website provides information about the quality control test, genotyping protocols, haplotype formats and informativity, population genetic analysis, literature references, and a list of contact addresses of the contributing laboratories.

A new method for the evaluation of matches in non-recombining genomes: application to Y-chromosomal short tandem repeat (STR) haplotypes in European males.

A 9-locus microsatellite framework (minimal haplotype), previously developed for forensic purposes so as to facilitate stain analysis, personal identification and kinship testing, has been adopted for the establishment of a large reference database of male European Y-chromosomal haplotypes. The extent of population stratification pertaining to this database, an issue crucial for its practical forensic application, was assessed through analysis of molecular variance (AMOVA) of the 20 regional samples included. Despite the notion of some significant haplotype frequency differences, which were found to correlate with known demographic and historic features of Europeans, AMOVA generally revealed a high level of genetic homogeneity among the populations analyzed. Owing to their high diversity, however, accurate frequency estimation is difficult for Y-STR haplotypes when realistic (i.e. moderately sized) datasets are being used. As expected, strong pair-wise and higher order allelic associations were found to exist between all markers studied, implying that haplotype frequencies cannot be estimated as products of allele frequencies. A new extrapolation method was therefore developed which treats haplotype frequencies as random variables and generates estimates of the underlying distribution functions on the basis of closely related haplotypes. This approach, termed frequency 'surveying', is based upon standard population genetics theory and can in principle be applied to any combination of markers located on the Y-chromosome or in the mitochondrial genome. Application of the method to the quality assured reference Y-STR haplotype database described herein will prove very useful for the evaluation of positive trace-donor matches in forensic casework.

[Examples for use of the STR systems in stain assessment with special reference to Y chromosome systems]. / Beispiele der Anwendung von STR-Systemen in der Spurenkunde unter besonderer Berücksichtigung von Y-chromosomalen Systemen.

The authors describe 3 cases where Y-chromosomal systems were used for typing the biological traces. In the first case, a murder, for the major amount of cell material found on a dish towel (and analysed two years after the crime) female persons were excluded for the system amelogenin and Y-chromosomal systems. A brother of the victim could not be excluded for autosomal STR-systems. Upon confrontation with the results of the DNA-analysis (among other things), this brother confessed the murder of his sister some days later. He was found guilty by the court. In the second case described, a rape of two girls, many traces were analysed parallely with Y-chromosomal and autosomal PCR-systems. The objects where male DNA matching the suspect were found (a paper tissue, a sweat shirt and the knickers of the girls), also showed small amounts of alleles matching with the suspect for autosomal systems, while the major part in these systems was from the girls. The suspect was sentenced to many years imprisonment. In the third case, a possible rape of a young woman, a stained microscope slide of a vaginal swab had to be examined. Microscopically a few sperm heads could be seen in a surplus of leucocytes. The male proportion could be analysed only in the Y-chromosomal systems, not in the autosomal ones. For the frequency calculation of the Y-chromosomal allele combination the haplotype data bank of the Institute for Legal Medicine of the Humboldt university in Berlin was indispensable.

Immunoelectron microscopic studies on the location of ribosomal proteins on the surface of the 40S ribosomal subunit from rat liver.

Seven ribosomal proteins have been localized by means of immunoelectron microscopy on the surface of the 40S ribosomal subunit from rat liver using monospecific antibodies. The location of ribosomal proteins S13/16, S19, and S24 is described for the first time, and that of ribosomal proteins S2, S3, S3a, and S7, which has been published previously on the basis of experiments performed with less well characterized antibody preparations [Lutsch et al., Mol. Gen. Genet. 176, 281-291 (1979) and Biomed. Biochim. Acta 42, 705-723 (1983)], is corrected in this paper. The results are discussed with respect to the involvement of these proteins in functional sites of the 40S ribosomal subunit.

Studies on interaction of 5 S RNA with ribosomal proteins.

Proteins of the large ribosomal subunit of rat liver (TP 60) were immobilized by diffusion transfer onto nitrocellulose after two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). Incubation of the TP 60 blots with 32P-labeled 5 S RNA under defined ionic conditions (300 mM KCl, 20 mM MgCl2) resulted in specific binding to a limited set of ribosomal proteins consisting of proteins L3, L4, L6, L13/15 and--to a lesser extent--L7 and L19. Under identical conditions, blots with proteins of the small ribosomal subunit (TP 40) did not bind 5 S RNA.

Preparation of 40S ribosomal subunit proteins of rat liver by combination of ion exchange chromatography with reversed phase liquid chromatography.

Total protein of small subunits of rat liver ribosomes was fractionated by chromatography on carboxymethylcellulose. Aliquots (between 5 and 30 mg) of the protein material of each peak were further separated by reversed phase liquid chromatography on an octadecasilyl silica column. In this way 15 of the 32 proteins of the small ribosomal subunit of rat liver could be isolated in milligram amounts.

Phosphorylation of ribosomal proteins as a possible control system for protein synthesis. Binding of Met-tRNAf to 40 S ribosomal subunits.

The evidence that protein S6 of rat liver ribosomes is involved in P-site functions and that this protein is the main target of the small subunit for in vivo phosphorylation suggests that S6 phosphorylation may contribute to the regulation of protein synthesis. Therefore, we have studied the activity of small ribosomal subunits with unphosphorylated and phosphorylated protein S6 in Met-tRNAf binding. The results described in this paper show that at least under in vitro conditions S6 phosphorylation does obviously not influence the activity of small ribosomal subunits for eIF-2 dependent binding of initiator-tRNA.

Action of rat liver cathepsin L on glucagon.

The proteolytic specificity of cathepsin L on glucagon was determined. Major cleavages are found between Thr7 and Ser8, Asp15 and Ser16, and between Met27 and Asn28. The bonds Ser11-Lys12, Val23-Gln24, and Gln24-Trp25 are hydrolyzed to a relatively low extent only. Whereas cathepsin B hydroxyzes glucagon at the C-terminus by a peptidyldipeptidase mechanism, cathepsin L cleaves the same substrate clearly as endopeptidase.