Biomedical subjects

G Klug

Publications and source records attributed to G Klug.

At least 37 records · Page 2Linked to original sources

PubMed1998

Integration host factor affects the oxygen-regulated expression of photosynthesis genes in Rhodobacter capsulatus.

The puf and puc operons of Rhodobacter capsulatus, which encode pigment binding proteins, are regulated by oxygen tension. Strain IR4 which carries a point mutation in the himA gene for integration host factor (IHF) shows a delayed increase in the amount of photosynthetic complexes after reduction of oxygen tension when compared to the isogenic wild-type strain. This delay in the formation of photosynthetic complexes correlates with a delay in increasing the level of transcription of the puf and puc operons, whereas the transcription of the bchD genes is not affected by IHF. Several IHF binding sites can be identified upstream of the puf and puc transcriptional starts by in vitro footprint analysis. Our results suggest that IHF facilitates the binding of other trans-acting factors, e.g. RegA, which in turn stimulate transcription of the puf and puc genes under low oxygen tension.

Bacterial Proteins↗

PubMed1998

Cloning and characterization of the rpoH gene of Rhodobacter capsulatus.

By using an oligonucleotide mixture corresponding to a region highly conserved among alternative sigma factors we identified a new sigma factor gene (rpoH) from Rhodobacter capsulatus. This gene encodes a protein of 34 kDa with strong similarity to the RpoH (sigma32) factors from other bacterial species. It was not possible to inactivate the R. capsulatus rpoH gene by introducing a resistance cassette, implying that it is essential for growth. The 5' ends of the mRNAs were mapped to two sequences with similarity to an rpoH- and an rpoD-dependent promoter, respectively. The amounts of both these mRNAs increased after heat shock, but were unaffected by a decrease in oxygen tension. Western analysis using a sigma factor-specific antibody revealed the accumulation of a protein of about 34 kDa after heat shock, and an increase in the amounts of a protein with the same size after reduction of oxygen tension in R. capsulatus cultures.

Amino Acid Sequence↗

PubMed1998

Prevalence, patterns and role of personality disorders in panic disorder patients with and without comorbid (lifetime) major depression.

A total of 84 consecutive out-patients from the Anxiety Disorders Clinic of the Psychiatric University Hospital in Graz with a current panic disorder were diagnosed for Axis I and II disorders using the Structured Clinical Interviews for DSM-III-R. The subjects were divided into two groups: (i) 49 patients who met the criteria for panic disorder with or without agoraphobia and had no history of an affective disorder and (ii) 35 patients who had a (lifetime) comorbidity of a major depressive disorder. There was a statistically significant difference in the prevalence of personality disorders between the two groups, which was due to the higher frequency of narcissistic personality disorder in the comorbid sample. Logistic regression analysis revealed that agoraphobia and/or major depression were associated with personality disorders, thus indicating that the relationship between panic disorder, agoraphobia and major depression is not straightforward, but is strongly influenced by the presence of Axis II disorders. Furthermore, the results of this study provide support for the 'unitary position' concerning the relationship between panic disorder and depression.

Adult↗

PubMed1998

Effect of oxygen on translation and posttranslational steps in expression of photosynthesis genes in Rhodobacter capsulatus.

The formation of the photosynthetic apparatus in Rhodobacter capsulatus is regulated by oxygen tension. Previous studies have shown a regulatory effect of oxygen on the transcription of photosynthesis genes and on the stability of certain mRNA segments. Here we show that oxygen affects puf and puc gene expression posttranslationally and that this regulation depends on the presence of bacteriochlorophyll. Our data suggest that this posttranslational effect of oxygen on puf and puc expression is due to the primary effect of oxygen on bacteriochlorophyll synthesis or assembly of pigment protein complexes. Oxygen does not affect the rates of translation of puf-encoded proteins.

Aerobiosis↗

PubMed1998

Molecular cloning and expression analysis of the Rhodobacter capsulatus sodB gene, encoding an iron superoxide dismutase.

Genetic complementation of a sodA sodB Escherichia coli mutant strain was used to clone Rhodobacter capsulatus genes involved in detoxification of superoxide radicals. After sequence analysis, 1 of the 16 identical clones obtained by this selection procedure was shown to contain an open reading frame with sequence similarity to that coding for Fe-containing superoxide dismutases (SodB). The R. capsulatus sodB gene was expressed in E. coli, and the nature of the metal ligand was confirmed by inhibitor sensitivity assays with lysates from both bacterial species. Activity staining of cleared Rhodobacter lysates resolved by polyacrylamide gel electrophoresis indicated that SodB was the only superoxide dismutase present in this phototrophic organism. The sodB gene was expressed at low levels in R. capsulatus cells grown under anaerobic or semiaerobic conditions, but expression was strongly induced upon exposure of the bacteria to air or to methyl viologen. Attempts to construct a sodB mutant in this organism by allelic exchange of the chromosomal copy of the gene with a suicide plasmid containing a mutated sodB gene were unsuccessful, strongly suggesting that the encoded superoxide dismutase is essential for viability of R. capsulatus in aerobic cultures.

Aerobiosis↗

PubMed1997

Identification, sequence analysis, and expression of the lepB gene for a leader peptidase in Rhodobacter capsulatus.

The leader peptidase (signal peptidase I) gene, lepB, of Rhodobacter capsulatus has been cloned and sequenced. The amino acid sequence of the predicted protein exhibits similarity to other known bacterial leader peptidases. R. capsulatus belongs to the alpha-subdivision of purple bacteria and thus is a relative of mitochondria in eukaryotes. Like the yeast mitochondrial inner membrane proteases IMP1 and IMP2, the leader peptidase from Rhodobacter has only one membrane-spanning segment. Sequence comparison of the Rhodobacter Lep protein with IMP1 and IMP2 did not reveal a higher overall similarity than between other prokaryotic signal peptidases and the mitochondrial enzymes. Expression studies using lacZ fusions in combination with primer extension analysis provide evidence for a weak promoter located a short distance from the transcription start of the lepB gene. Failure to establish a Rhodobacter strain with a disrupted lepB gene indicates that this gene is essential.

Amino Acid Sequence↗

PubMed1997

[Psychiatric aspects in rehabilitation patients and elderly patients].

From a psychiatric point of view reintegration and integration meaning a reorientation of the individual should be the points to be stressed in respect of rehabilitation of elder people. These individuals should be led to finding new positions in their psychosocial context. It would be an easier task to reach having developed some crucial social abilities in earlier stages of life. These abilities should be established already. We want to demonstrate this considering individual activities, social abilities, trusting and flexibility. Getting older is a developmental process. You cannot develop abilities which you never were confronted with before. If you do not seed you will not get a crop. Depending on early set resources individuals will later be able to integrate, reintegrate or not. This is to be set in an early stage of life.

Activities of Daily Living↗

PubMed1997

Predicting recovery from head injury in young children: a prospective analysis.

It has been argued that young children's brains are "plastic," and may sustain substantial brain insult with little loss of function. Recent research suggests that this notion may not apply for generalized cerebral pathology. The present study aimed to evaluate this proposition using a sample of 73 young children, divided into 3 groups: severe head injury (HI; N = 17); mild-moderate HI (N = 32); and noninjured controls (N = 24). Preinjury screening established equivalence across groups for age, sex, preinjury ability, behavioral adjustment, socioeconomic status, and family functioning. Children were evaluated as soon as possible postinjury, and again 12 months postinjury, in three domains: intellectual ability, language, and memory. Results indicated that severe HI was associated with substantial, persisting difficulties in all areas. In contrast, children with mild-moderate HI experienced fewer difficulties, and often performed similarly to controls, both acutely and 12 months postinjury. There was no evidence of differential recovery of function associated with injury severity, with performance increments consistent across groups and probably due to either age-appropriate developmental gains, or test-related practice effects. Poorer outcome at 12 months postinjury was predicted by injury severity primarily, with earlier age at injury, and premorbid ability associated with outcome in specific domains.

Age Factors↗

PubMed1997

Predictors and indicators of academic outcome in children 2 years following traumatic brain injury.

Within the context of a longitudinal study investigating outcome for children following traumatic brain injury, this paper reports on the utility of neuropsychological testing in predicting academic outcome in children 2 years following traumatic brain injury (TBI). Twenty-nine school-age children who were admitted to hospital after TBI were assessed with a battery of neuropsychological and academic measures at 3 and 24 months postinjury. The neuropsychological battery included measures of memory, learning, and speed of information processing. Academic outcome was assessed in terms of post-TBI change in school placement. According to logistic regression analysis, change in placement from regular to special education at 2 years post-TBI was predicted by injury severity and by neuropsychological performance at 3 months post-TBI. Findings suggest that neuropsychological testing is useful in identifying children with special educational needs subsequent to TBI.

Adolescent↗

PubMed1996

Cloning, nucleotide sequence and characterization of the rpoD gene encoding the primary sigma factor of Rhodobacter capsulatus.

A synthetic oligodeoxynucleotide probe based on a highly conserved region of the sigma factors was used to identify and clone the rpoD gene encoding the principal sigma factor of R. capsulatus. The deduced polypeptide contains 674 amino acids and has a predicted molecular mass of 75,942 Da. The deduced amino acid sequence of R. capsulatus RpoD protein exhibits 46.2% and 45.7% identity with housekeeping sigma factors of Pseudomonas and E. coli, respectively. Unsuccessful attempts to inactivate the single chromosomal rpoD gene of R. capsulatus by gene replacement technique indicate that this gene is essential for cell survival, as expected for the primary sigma factor. The rpoD transcript 5'-end was mapped by primer extension analysis, 74 bp upstream of the initiation codon and DNA sequence analysis has identified a motif resembling the delta 70 E. coli consensus promoter sequences at the expected distance from this proposed transcription start site. rpoD gene expression, as measured by the activity of the phi (rpoD'-lacZ+) translational fusion, was found to be constant throughout exponential and early plateau phases, but significantly increased at later times of the stationary phase.

Amino Acid Sequence↗

PubMed1996

Identification and analysis of the rnc gene for RNase III in Rhodobacter capsulatus.

The large subunit ribosomal RNA of the purple bacterium Rhodobacter capsulatus shows fragmentation into pieces of 14 and 16S, both fragments forming the functional equivalent of intact 23S rRNA. An RNA-processing step removes an extra stem-loop structure from the 23S rRNA [Kordes, E., Jock, S., Fritsch, J., Bosch, F. and Klug, G. (1994) J. Bacteriol., 176, 1121-1127]. Taking advantage of the fragmentation deficient mutant strain Fm65, we used genetic complementation to find the mutated gene responsible for this aberration. It was identified as the Rhodobacter homologue to mc from Escherichia coli encoding endoribonuclease III (RNase III). The predicted protein has 226 amino acids with a molecular weight of 25.5 kDa. It shares high homology with other known RNase III enzymes over the full length. In particular it shows the double-stranded RNA-binding domain (dsRBD) motif essential for binding of dsRNA substrates. The Fm65 mutant has a frame shift mutation resulting in complete loss of the dsRBD rendering the enzyme inactive. The cloned Rhodobacter enzyme can substitute RNase III activity in an RNase III deficient E. coli strain. Contrary to E. coli, the Rhodobacter mc is in one operon together with the lep gene encoding the leader peptidase.

Amino Acid Sequence↗

PubMed1996

Expression of the thioredoxin gene (trxA) in Rhodobacter sphaeroides Y is regulated by oxygen.

The structural gene (trxA) coding for thioredoxin in the photosynthetic bacterium Rhodobacter sphaeroides has been cloned and sequenced previously. In the present study, the role of oxygen in trxA expression in R. sphaeroides Y was investigated using mRNA analyses and plasmid-borne trxA'-lacZ+ translational and transcriptional fusions. Northern analysis revealed a trxA-specific transcript of approximately 420-460 nucleotides, indicating that trxA is transcribed as a single gene. By studying the beta-galactosidase activity in strains harboring various phi(trxA'-lacZ+) fusion constructs, the promoter region of the trxA gene was localized within a 64-bp region located 97 nucleotides upstream of the trxA initiator codon. A single trxA transcription initiation site was mapped by primer extension, 27 bp upstream of the trxA gene. Based on these results and the DNA sequence analysis, we propose that a sigma70 consensus sequence serves as a trxA promoter. Results from oxygen shift experiments, as deduced from both mRNA analysis and fusions of the trxA promoter region to lacZ indicate that transcription of the R. sphaeroides trxA gene is regulated by high oxygen tension. DNA sequences involved in this oxygen regulation were also localized in the 64-bp region containing the trxA promoter. Based on our findings the hypothetical biological function of thioredoxin from R. sphaeroides is discussed.

Base Sequence↗

PubMed1996

Effect of the pufQ-pufB intercistronic region on puf mRNA stability in Rhodobacter capsulatus.

Differential expression of genes localized within the polycistronic puf operon of Rhodobacter capsulatus is partly due to altered stabilities of individual mRNA segments. We show that the 5' untranslated region (UTR) of pufB contributes to the unusual longevity of the 0.5 kb light-harvesting (LH) I specific pufBA mRNA and of the 2.7 kb pufBALMX mRNA. Three stem-loop structures have been identified within the pufQ-pufB intercistronic region by means of RNA secondary-structure analysis in vitro and in vivo. Deletion analysis of the pufB 5' UTR indicates that the complete set of secondary structures is required to maintain wild-type levels of pufBA mRNA stability. A phylogenetic comparison of pufB 5' UTRs of other photosynthetic bacteria reveals an evolutionary conservation of the base-pairing potential despite sequence divergence. Comparison of puf mRNA decay in Escherichia coli strains with or without endoribonuclease E (RNase E) activity suggests that the pufB 5' secondary structures protect the downstream mRNA segment against degradation by RNase E. Removal of the 117-nucleotide pufQ-pufB intercistronic region results in loss of stability for the pufBA and pufBALMX mRNAs with concomitant stabilization of the full-length puf primary transcript (QBALMX). We therefore conclude that the deleted sequence functions both as a stabilizing element for pufBALMX and pufBA segments and as a target site for initial rate-limiting decay of the unstable pufQBALMX mRNA.

Bacterial Proteins↗

PubMed1996

The bluF gene of Rhodobacter capsulatus is involved in conversion of cobinamide to cobalamin (vitamin B12).

The bluF gene of Rhodobacter capsulatus is the first gene of the bluFEDCB operon which is involved in late steps of the cobalamin synthesis. To determine the function of the bluF gene product, a bluF::omega-Km mutant strain was constructed and characterized. This vitamin B12 auxotrophic mutant strain shows a 3.5-times higher vitamin B12 requirement under phototrophic growth conditions than under chemotrophic growth conditions. Surprisingly, the bluF promoter activity does not respond to alterations to the oxygen tension or vitamin B12 concentration.

Bacterial Proteins↗

PubMed1995

Neuropsychological deficit and academic performance in children and adolescents following traumatic brain injury.

Evaluated the utility of neuropsychological testing in predicting academic outcome in children 1 year following traumatic brain injury (TBI). Fifty-one school age children who were admitted to hospital after TBI were assessed with a battery of neuropsychological measures at 3 months postinjury. Academic achievement was assessed at 3 and 12 months postinjury. The neuropsychological battery included intelligence testing and measures of memory, learning, and speed of information processing. Academic outcome was assessed in terms of post-TBI changes in reading, spelling, and arithmetic; changes in teacher ratings of school performance; and change in school placement. According to logistic regression analysis, change in placement from regular to special education at 1-year post-TBI was predicted by injury severity and by neuropsychological performance at 3 months post-TBI. Findings suggest that neuropsychological testing is useful in identifying children with special educational needs subsequent to TBI.

Achievement↗

PubMed1995

Identification of an mRNA element promoting rate-limiting cleavage of the polycistronic puf mRNA in Rhodobacter capsulatus by an enzyme similar to RNase E.

We have identified an mRNA element that is involved in the initial cleavage of the pufBALMX mRNA species in Rhodobacter capsulatus. This endoribonuclease recognition site, the first to be identified in a bacterial species other than Escherichia coli, shows strong similarities to mRNA sequences cleaved by the endoribonuclease E in E. coli. The presence of an RNase E-like enzyme in R. capsulatus is further supported by in vitro cleavage of E. coli transcripts by R. capsulatus extracts at sites attributed to RNase E and by the cross-reaction of a polypeptide from R. capsulatus with antisera against E. coli RNase E. Our data provide evidence that mRNAs are degraded in different bacterial species by enzymes with similar recognition sequences and activities. We present a model that attributes the segmental differences in stability of the polycistronic puf transcript to a specific distribution of mRNA decay-promoting and mRNA decay-impeding elements.

Base Sequence↗

PubMed1995

Identification and sequence analysis of genes involved in late steps in cobalamin (vitamin B12) synthesis in Rhodobacter capsulatus.

A 6.4-kb region of a 6.8-kb BamHI fragment carrying Rhodobacter capsulatus genes involved in late steps of cobalamin synthesis has been sequenced. The nucleotide sequence and genetic analysis revealed that this fragment contains eight genes arranged in at least three operons. Five of these eight genes show homology to genes involved in the cobalamin synthesis of Pseudomonas denitrificans and Salmonella typhimurium. The arrangement of these homologous genes differs considerably in the three genera. Upstream of five overlapping genes (named bluFEDCB), a promoter activity could be detected by using lacZ fusions. This promoter shows no regulation by oxygen, vitamin B12 (cobalamin), or cobinamide. Disruption of the bluE gene by a Tn5 insertion (strain AH2) results in reduced expression of the puf and puc operons, which encode pigment-binding proteins of the photosynthetic apparatus. The mutant strain AH2 can be corrected to a wild-type-like phenotype by addition of vitamin B12 or cobinamide dicyanide. Disruption of the bluB gene by an interposon (strain BB1) also disturbs the formation of the photosynthetic apparatus. The mutation of strain BB1 can be corrected by vitamin B12 but not by cobinamide. We propose that a lack of cobalamin results in deregulation and a decreased formation of the photosynthetic apparatus.

Amino Acid Sequence↗

PubMed1994

Cloning of a gene involved in rRNA precursor processing and 23S rRNA cleavage in Rhodobacter capsulatus.

In Rhodobacter capsulatus wild-type strains, the 23S rRNA is cleaved into [16S] and [14S] rRNA molecules. Our data show that a region predicted to form a hairpin-loop structure is removed from the 23S rRNA during this processing step. We have analyzed the processing of rRNA in the wild type and in the mutant strain Fm65, which does not cleave the 23S rRNA. In addition to the lack of 23S rRNA processing, strain Fm65 shows impeded processing of a larger 5.6-kb rRNA precursor and slow maturation of 23S and 16S rRNAs from pre-23S and pre-16S rRNA species. Similar effects have also been described previously for Escherichia coli RNase III mutants. Processing of the 5.6-kb precursor was independent of protein synthesis, while the cleavage of 23S rRNA to generate 16S and 14S rRNA required protein synthesis. We identified a DNA fragment of the wild-type R. capsulatus chromosome that conferred normal processing of 5.6-kb rRNA and 23S rRNA when it was expressed in strain Fm65.

Bacteriochlorophylls↗