Forgot your password?
New user?
New Window Opens on the Secret Life of Microbes: Scientists Develop First Microbial Profiles of Ecosystems
CAS announces the release of SciFinder Scholar for Mac OS X
Press Releases
Press Releases
| Check out our New Publishers' Select for Free Articles |
Archives of Microbiology (v.167, #4)
The [NiFe] hydrogenases of Methanococcus voltae: genes, enzymes and regulation by Oliver Sorgenfrei; Steffen Müller; Matthias Pfeiffer; Izabela Sniezko; A. Klein (pp. 189-195).
Methanococcus voltae carries genetic information for four [NiFe] hydrogenases. Two of the hydrogenases are predicted to contain selenocysteine on the basis of in-frame TGA codons, while the genes encoding the two other enzymes contain cysteine codons at homologous positions. Their predicted subunit compositions and their electron acceptor specificities are similar to those of the respective selenium-containing enzymes. The selenium-containing hydrogenases have been purified and characterized. Only one of them reduces the deazaflavin F420. The activity of the F420-nonreducing enzyme is exceptionally high. The selenium atom has been shown by EPR spectroscopy to be a ligand to the Ni atom in the primary reaction centers in both enzymes. The spectroscopic analyses also yielded a description of the electronic configuration around the NiFe center at different oxidation states and in the presence of the competitive inhibitor, CO. The genes encoding the selenium-free hydrogenases are expressed only in the absence of selenium. They are linked by an intergenic region in which regulatory cis elements were defined by employing reporter gene constructs and site-directed mutagenesis.
Keywords: Key words [NiFe] Hydrogenases; Methanococcus; voltae; Archaea; Selenocysteine; Selenium-dependent; gene regulation; Operator; Silencer; EPR
A deletion in the sapA homologue cluster is responsible for the loss of the S-layer in Campylobacter fetus strain TK by Masaki Fujita; Tetsuhiro Moriya; S. Fujimoto; Nobuyuki Hara; Kazunobu Amako (pp. 196-201).
The surface array protein (SAP) of Campylobacter fetus strain TK is encoded by seven homologous sapA genes clustered on the chromosomal DNA. The spontaneously arising variant strain TK(SAP–) produces no SAP and carries an approximately 10-kb chromosomal deletion. To elucidate the mechanism underlying the loss of SAP synthesis, we analyzed the region containing the sapA homologues and the deletion. We constructed a physical map of the sapA cluster region by aligning the clones that contain sapA homologues. These analyses demonstrated that all sapA homologues were located within a limited region of about 50 kb of chromosomal DNA of strain TK. The TK(SAP–) deletion was located within this cluster and was 13.3 kb in size. The deletion occurred between two sapA homologues and resulted in the formation of a chimeric sapA homologue in the variant strain. Sequence analysis of the upstream regions and the conserved regions of all sapA homologues revealed a high degree of similarity. However, only one sapA homologue contained a putative promoter sequence. This promoter sequence was located in the deleted region. Thus, the deletion of the promoter appears to be responsible for the loss of SAP expression in TK(SAP–).
Keywords: Key wordsCampylobacter fetus; sapA Homologues; Promoter; Chi sequence; Antigenic variation; Surface layer
Molecular cloning of the neutral trehalase gene from Kluyveromyces lactis and the distinction between neutral and acid trehalases by F. C. Amaral; P. Van Dijck; J. R. Nicoli; J. M. Thevelein (pp. 202-208).
We cloned the Kluyveromyces lactis KlNTH1 gene, which encodes neutral trehalase. It showed 65.2% and 68.5% identity at nucleotide and amino acid sequence level, respectively, with the Saccharomyces cerevisiae NTH1 gene. Multiple alignment of the predicted trehalase protein sequences from yeasts, bacteria, insects, and mammals revealed two major domains of conservation. Only the yeast trehalases displayed in an N-terminal extension two consensus sites for cAMP-dependent protein phosphorylation and a putative Ca2+-binding sequence. Gene disruption of the KlNTH1 gene abolished neutral trehalase activity and clearly revealed a trehalase activity with an acid pH optimum. It also resulted in a high constitutive trehalose level. Expression of the KlNTH1 gene in an S. cerevisiae nth1Δ mutant resulted in rapid activation of the heterologous trehalase upon addition of glucose to cells growing on a nonfermentable carbon source and upon addition of a nitrogen source to cells starved for nitrogen in a glucose-containing medium. In K. lactis, the same responses were observed except that rapid activation by glucose was observed only in early-exponential-phase cells. Inactivation of K. lactis neutral trehalase by alkaline phosphatase and activation by cAMP in cell extracts are consistent with control of the enzyme by cAMP-dependent protein phosphorylation.
Keywords: Key words Trehalase; Trehalose; Kluyveromyces lactis; Protein phosphorylation; NTH1; Glucose induction; Glucose transport
Effect of chelating agents and respiratory inhibitors on regulation of the cadA gene in Escherichia coli by S. G. Reams; Norizan Lee; F. Mat-Jan; D. P. Clark (pp. 209-216).
The cadA gene that encodes lysine decarboxylase in Escherichia coli is induced by low pH and – during anaerobic growth – by the substrate, lysine. We used operon fusions of cadA to lacZ to investigate the effects of aeration on cadA regulation. When an insertion mutation in osmZ (= hns) was introduced, a cadA-lacZ fusion was derepressed in the presence of air to approximately the same level as seen during anaerobic growth. However, the pH-dependent regulation of cadA was not affected by osmZ. Introduction of mutations in rpoS, fur, or fnr had no significant effect on cadA expression. However, defects in arcB or arcA largely abolished expression of cadA during anaerobic growth. Nonetheless, strains defective in both arcB and osmZ showed the same high cadA-lac expression in air as seen in the single osmZ derivatives. Blocking the respiratory chain with mutations or chemical inhibitors also caused derepression of a cadA-lacZ fusion in air, while agents affecting the proton gradient had no effect. Derepression of cadA in air was also mediated by several chelating agents, in particular by methoxyindole carboxylic acid. Addition of Fe2+ overcame this effect. Chelating agents also abolished the expression during aerobic growth of several genes known to be under arcAB control and which are normally repressed during anaerobic growth but induced in the presence of air. This implies that the effect of chelating agents on cadA expression is mediated via the arcAB regulatory system.
Keywords: Key words Anaerobic metabolism; Lysine; decarboxylase; β-Galactosidase; Gene fusions; Cyanide; Respiratory inhibitors; Arc regulation; Chelating agents
Biochemical and phylogenetic characterization of two novel deep-sea Thermococcus isolates with potentially biotechnological applications by F. Canganella; W. Jack Jones; Agata Gambacorta; Garabed Antranikian (pp. 233-238).
The partial 16S rDNA gene sequences of two thermophilic archaeal strains, TY and TYS, previously isolated from the Guaymas Basin hydrothermal vent site were determined. Lipid analyses and a comparative analysis performed with 16S rDNA sequences of similar thermophilic species showed that the strains isolated from deep-sea vents were not identical to the other species belonging to the genus Thermococcus. On the basis of the results of the phylogenetic analyses, lipid analyses, and previously reported physiological data, we believe that strains TY and TYS are significantly different from the previously described Thermococcus species. According to specific physiological and molecular features, we propose the use of these isolates as potential tools for the development of biotechnological applications in the field of starch processing and DNA technology.
Keywords: Key words Deep-sea hydrothermal vents; Thermophilic archaea; Thermococcus; Biotechnological applications
Two alternative mechanisms of cell separation in staphylococci: one lytic and one mechanical by P. Giesbrecht; Thomas Kersten; Heinrich Maidhof; Jörg Wecke (pp. 239-250).
Electron microscopy studies revealed two different mechanisms of cell separation in Staphylococcus aureus. Both mechanisms were initiated by the centrifugal lytic action (directed outward from the center) of murosomes, which perforated the peripheral cell wall. Thereafter, during the first type of cell separation, murosomes also lysed large parts of the cross wall proper in the opposite, i.e., centripetal direction, forming spokelike lytic lesions (“separation scars”) next to the most prominent structure of the cross wall, the splitting system. This bidirectional lytic action of murosomes revealed that the staphylococcal cross wall is composed of permanent and transitory parts; transitory parts constituted about one-third of the volume of the total cross wall and seemed to be digested during cell separation. The second mechanism of cell separation was encountered within the splitting system, which has been regarded as the main control unit for lytic cell separation for more than 25 years. The splitting system, however, represents mainly a mechanical aid for cell separation and becomes effective when cell-wall autolytic activities are insufficient.
Keywords: Key wordsStaphylococcus aureus; Cell wall; Morphogenesis; Cell separation; Murosomes; Penicillin-splitting system; Wall autolysins
A polyketide-synthase-like gene is involved in the synthesis of heterocyst glycolipids in Nostoc punctiforme strain ATCC 29133 by Elsie L. Campbell; Michael F. Cohen; J. C. Meeks (pp. 251-258).
A Tn5-1063-derived mutant of Nostoc punctiforme strain ATCC 29133 was unable to fix N2 in air although it reduced acetylene in the absence of O2. Mutant strain UCD 307 formed cells morphologically similar to heterocysts, but it failed to synthesize the characteristic heterocyst glycolipids. Sequence analysis around the site of insertion revealed an ORF of 3,159 base pairs, termed hglE. hglE putatively encodes a 115.4-kDa protein containing two domains with conserved amino acid sequences identified with acyl transferase and the chain length factor variation of β-ketoacyl synthase active sites. These active sites are characteristic of polyketide and fatty acid synthases. The N. punctiforme strain 29133 hglE gene is transcribed only under nitrogen-limiting growth conditions. The hglE gene, or similar sequences, was found in all other heterocyst-forming cyanobacteria surveyed and was absent in unicellular Synechococcus sp. strain PCC 7942. Based on these results, we propose that the synthesis of heterocyst glycolipids follows a pathway characteristic of polyketide synthesis and involves similar large, multienzyme complexes.
Keywords: Key words Fox; mutant; Heterocyst glycolipid; hglE; Nostoc punctiforme; Polyketide synthase