Promoter characterization and genomic organization of the gene encoding integrin-linked kinase 1

in Biochimica et Biophysica Acta-Gene Structure and Expression (2002), 1575(1-3), 117-122

Integrin-linked kinase (ILK)-1 is a 59-kDa serine-threonine protein kinase, which associates with the cytoplasmic domain of β1, β2 and β3 integrins and acts as a receptor proximal kinase regulating ... [more ▼]

Integrin-linked kinase (ILK)-1 is a 59-kDa serine-threonine protein kinase, which associates with the cytoplasmic domain of β1, β2 and β3 integrins and acts as a receptor proximal kinase regulating integrin-mediated signal transduction. We have recently identified an isoform of ILK (ILK-2), which is expressed, in a TGF-β1-dependent manner, in a highly invasive tumor cell line but not in normal adult tissues. In contrast, ILK-1 is ubiquitously expressed in normal tissues and is up-regulated in various tumors independent of TGF-β1. Here, we report the structural organization and the promoter activity of the human ILK-1 gene, contained within a 8.8-kb genomic fragment cloned from a human BAC library. The mature protein is encoded by 13 exons. The last coding exon contains the entire 3′ UTR of the ILK-1 gene, which overlaps with the complementary 3′ UTR sequence of the TAF2H gene, a TATA box binding protein-associated factor. A major transcriptional initiation start site was found 138 bp upstream of exon 1 in close proximity to a consensus initiator element (Inr). The ILK gene is transcribed by a TATA-less and CAAT-less promoter with typical features of housekeeping genes. The promoter activity was characterized by a luciferase reporter assay and the minimal sequence conferring promoter activity was 349 bp in size and located immediately upstream of exon 1. © 2002 Elsevier Science B.V. All rights reserved. [less ▲]

Placental transfer and decay of maternally acquired antimeasles antibodies in Nigerian children

in Pediatric Infectious Disease Journal (2000), 19(7), 635-641

Background. In developing countries vaccination against measles virus (MV) is generally administered at 9 months of age, although it is well- documented that protection of most infants by passively ... [more ▼]

Background. In developing countries vaccination against measles virus (MV) is generally administered at 9 months of age, although it is well- documented that protection of most infants by passively acquired maternal MV antibodies is waning before immunization is given. The purpose of this study was to investigate the decay of maternally derived MV antibodies in Nigerian infants as well as to compare a German and Nigerian cohort of paired mothers and newborns regarding the placental transfer efficiency of MV-specific IgG and total IgG antibodies. Methods. MV-specific IgG antibodies were measured with a commercially available MV-enzyme-linked immunosorbent assay, a recombinant hemagglutinin enzyme-linked immunosorbent assay as well as a neutralization assay. Total IgG values were determined with a standard immunoturbidimetric test. Results. Anti-MV IgG titers were twice as high in German newborns as in Nigerian newborns. An increased concentration of immunoglobulins transferred via the placenta was found only in the German cohort. High concentrations of total maternal IgG reduced the concentration of MV-specific as well as total IgG that crossed the placenta. Furthermore only 17% of the 4-month-old Nigerian infants were still protected against measles. Antibodies had a biologic half-life of 33 days and a biochemical half-life of 48 days. Conclusions. Our findings demonstrate that the decay of passively acquired MV antibodies occurred even more rapidly than expected resulting in susceptibility to MV in most of the 4-month-old infants in Nigeria. Furthermore transfer of maternal anti-MV IgG and total IgG antibodies to the newborn was more efficient in the German cohort compared with the Nigerian group. These findings suggest the use of alternative vaccination strategies in developing countries to possibly reduce the window of susceptibility against measles. [less ▲]

Genotypic and antigenic characterization of hemagglutinin proteins of African measles virus isolates

in Virus Research (1999), 62(1), 89-95

A comprehensive phylogenetic study based on the hemagglutinin (H) protein of all known African measles virus (MV) isolates is presented. The study includes 64 new H gene sequences from Ghana, Nigeria and ... [more ▼]

A comprehensive phylogenetic study based on the hemagglutinin (H) protein of all known African measles virus (MV) isolates is presented. The study includes 64 new H gene sequences from Ghana, Nigeria and South Africa as well as viruses from Zambia and The Gambia for which only incomplete sequencing data were available and that have previously not been genotyped. The results provide further support to the tentative assignment of the Nigerian and Ghanaian viruses to a new genotype B3 within clade B. A distinct geographic distribution pattern emerged with clade B viruses circulating exclusively in African countries north of the equator. All MV strains from southern Africa grouped in clades A and D with the majority of viruses belonging to genotype D4. The viruses considerably differed by their sensitivity to neutralization by monoclonal antibodies (mAb), but three selected antibodies were sufficient to distinguish between African MVs representing four different genotypes. Copyright (C) 1999 Elsevier Science B.V. [less ▲]

Partial amplification of the measles virus nucleocapsid gene from stored sera and cerebrospinal fluids for molecular epidemiological studies

in Journal of Medical Virology (1998), 56(2), 174-177

The analysis of stored sera for retrospective molecular epidemiological studies provides a powerful tool to investigate strain variation in measles viruses that had circulated up to 20 years ago. For this ... [more ▼]

The analysis of stored sera for retrospective molecular epidemiological studies provides a powerful tool to investigate strain variation in measles viruses that had circulated up to 20 years ago. For this purpose, a rapid and simple method for extraction of RNA from stored sera and cerebrospinal fluids (CSF) was developed. When used on sera and CSFs that have been frozen for as long as 20 years, this method proved to be more efficient than established techniques. The extracted RNA was reverse transcribed into cDNA by using random hexamer primers. The PCR amplification of the 3' terminus of the nucleocapsid gene (N) was divided into two overlapping fragments of 375 and 384 bp length, covering the entire region of interest. This region is thought to have the highest variability within the MV genome and has previously been shown to be suitable for strain characterization. The resulting PCR fragments were sequenced manually by using standard methods without the need of further clean-up steps. [less ▲]

Rapid identification of measles virus strains by the heteroduplex mobility assay

in Virus Research (1997), 47(2), 472197-203

The continued endemic presence of measles virus (MV), and the large number of isolates which are made in South Africa each year, demanded the use of a rapid and reliable pre-screening technique to select ... [more ▼]

The continued endemic presence of measles virus (MV), and the large number of isolates which are made in South Africa each year, demanded the use of a rapid and reliable pre-screening technique to select isolates for molecular epidemiological studies by sequence analysis. The heteroduplex mobility assay (HMA) was used to genetically characterize 47 MV isolates collected from three different provinces in South Africa, made between 1986 and 1995. The carboxyl-terminal 590 nt of the nucleocapsid (N) gene - the most variable region of the genome - was amplified by polymerase chain reaction (PCR) and subsequently subjected to HMA analysis for initial genotyping. The results showed three different patterns of heteroduplex formation by gel electrophoresis, representing two distinct wild-type lineages and one group of vaccine-like viruses. Comparison of HMA results with phylogenetic analysis of sequence data for several of the South African MV strains showed a complete correlation of results. The HMA proved to be a useful tool for screening MV isolates for use in molecular epidemiological studies. [less ▲]