AIM: To develop novel DNA extraction and typing procedure for DNA identification of the 7th century human remains, determine the familiar relationship between the individuals, estimate the Y-chromosome haplogroup, and compare the Y-chromosome haplotype with the contemporary populations. METHODS: DNA from preserved femur samples was extracted using the modified silica-based extraction technique. Polymerase chain reaction amplification was performed using human identification kits MiniFiler, Identifiler, and Y-filer and also laboratory-developed and validated Y-chromosome short tandem repeat (STR) pentaplexes with short amplicons. RESULTS: For 244A, 244B, 244C samples, full autosomal DNA profiles (15 STR markers and Amelogenin) and for 244D, 244E, 244F samples, MiniFiler profiles were produced. Y-chromosome haplotypes consisting of up to 24 STR markers were determined and used to predict the Y-chromosome haplogroups and compare the resulting haplotypes with the current population. Samples 244A, 244B, 244C, and 244D belong to Y-chromosome haplogroup R1b and the samples 244E and 244F to haplogroup G2a. Comparison of ancient haplotypes with the current population yielded numerous close matches with genetic distance below 2. CONCLUSION: Application of forensic genetics in archaeology enables retrieving new types of information and helps in data interpretation. The number of successfully typed autosomal and Y-STR loci from ancient specimens in this study is one of the largest published so far for aged samples.
- MeSH
- dějiny starověku MeSH
- DNA fingerprinting metody MeSH
- genetické markery MeSH
- genotyp MeSH
- geny vázané na chromozom Y MeSH
- lidé MeSH
- mikrosatelitní repetice MeSH
- polymerázová řetězová reakce MeSH
- soudní antropologie MeSH
- soudní genetika MeSH
- zkameněliny MeSH
- Check Tag
- dějiny starověku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- historické články MeSH
- práce podpořená grantem MeSH
Streptococcus pneumoniae carries a single Ser/Thr protein kinase gene stkP in its genome. Biochemical studies performed with recombinant StkP have revealed that this protein is a functional membrane-linked eukaryotic-type Ser/Thr protein kinase. Here, we demonstrate that the deletion of its extracellular domain negatively affects the stability of a core kinase domain. In contrast, the membrane anchored kinase domain and the full-length form of StkP were stable and capable of autophosphorylation. Furthermore, evidence is presented that StkP forms dimers through its transmembrane and extracellular domains.
Signal transduction pathways in both prokaryotes and eukaryotes utilize protein phosphorylation as a key regulatory mechanism. Recent studies have proven that eukaryotic-type serine/threonine protein kinases (Hank's type) are widespread in many bacteria, although little is known regarding the cellular processes they control. In this study, we have attempted to establish the role of a single eukaryotic-type protein kinase, StkP of Streptococcus pneumoniae, in bacterial survival. Our results indicate that the expression of StkP is important for the resistance of S. pneumoniae to various stress conditions. To investigate the impact of StkP on this phenotype, we compared the whole-genome expression profiles of the wild-type and DeltastkP mutant strains by microarray technology. This analysis revealed that StkP positively controls the transcription of a set of genes encoding functions involved in cell wall metabolism, pyrimidine biosynthesis, DNA repair, iron uptake, and oxidative stress response. Despite the reduced transformability of the stkP mutant, we found that the competence regulon was derepressed in the stkP mutant under conditions that normally repress natural competence development. Furthermore, the competence regulon was expressed independently of exogenous competence-stimulating peptide. In summary, our studies show that a eukaryotic-type serine/threonine protein kinase functions as a global regulator of gene expression in S. pneumoniae.
- MeSH
- bakteriální proteiny genetika metabolismus MeSH
- delece genu MeSH
- eukaryotické buňky enzymologie MeSH
- fenotyp MeSH
- financování organizované MeSH
- koncentrace vodíkových iontů MeSH
- mikrobiální viabilita genetika účinky léků MeSH
- mutace MeSH
- osmotický tlak MeSH
- oxidační stres MeSH
- peroxid vodíku farmakologie MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- protein-serin-threoninkinasy genetika metabolismus MeSH
- regulace genové exprese u bakterií MeSH
- sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
- Streptococcus pneumoniae enzymologie genetika růst a vývoj MeSH
- testy genetické komplementace MeSH
- vysoká teplota MeSH