Glyceroglycolipids are very important in Gram-positive bacteria and cyanobacteria. In Bacillus subtilis, a model organism for the Gram-positive bacteria, the ugtP mutant, which lacks glyceroglucolipids, shows abnormal morphology. Lack of glucolipids has many consequences: abnormal localization of the cytoskeletal protein MreB and activation of some extracytoplasmic function (ECF) sigma factors (σM, σV and σX) in the log phase are two examples. Conversely, the expression of monoglucosyldiacylglycerol (MGlcDG) by 1,2-diacylglycerol 3-glucosyltransferase from Acholeplasma laidlawii (alMGS) almost completely suppresses the ugtP disruptant phenotype. Activation of ECF sigmas in the ugtP mutant is decreased by alMGS expression, and is suppressed to low levels by MgSO4 addition. When alMGS and alDGS (A. laidlawii 1,2-diacylglycerol-3-glucose (1-2)-glucosyltransferase producing diglucosyldiacylglycerol (DGlcDG)) are simultaneously expressed, σX activation is repressed to wild type level. These observations suggest that MGlcDG molecules are required for maintenance of B. subtilis cell shape and regulation of ECF sigmas, and that DGlcDG regulates σX activity. The activation of ECF sigmas is not accompanied by proteolysis of anti-σ. Thus, glyceroglucolipids may have the specific role of helping membrane proteins function by acting in the manner of chaperones.
Proteins belonging to the sigma factor family in eubacteria initiate transcription by associating with RNA polymerase. A subfamily, the extracytoplasmic function (ECF) sigma factors, which form a widely distributed bacterial signal transduction system comprising a sigma factor and a cognate membrane-embedded anti-sigma factor, regulates genes in response to stressors that threaten cell envelope integrity including the cell wall and membrane. The Gram-positive soil bacterium Bacillussubtilis provides a valuable model for investigation of the ECF sigma factors. This review focuses on the function and regulation of ECF sigma factors in B. subtilis, in which anti-sigma factors play a role in connecting an external stimulus with gene regulation. As representative examples, the regulon and regulatory mechanism of σW are closely associated with membrane-active stressors, whereas σM is strongly induced by conditions that impair peptidoglycan synthesis. These studies demonstrate that the mechanisms of ECF-dependent signaling are divergent and constitute a multi-layered hierarchy, and provide useful insights into the elucidation of unknown mechanisms related to ECF sigma factors.
Glycolipids constitute the majority of membrane components in oxygenic photosynthetic organisms, whereas they are minor lipids in other organisms. In cyanobacteria, three glycolipids comprise ~90 mol% of the total lipids in thylakoid membranes, where photosynthetic electron transport occurs. Among these glycolipids, 80 mol% are galactolipids (monogalactosyldiacylglycerol and digalactosyldiacylglycerol). Galactolipids are well conserved in oxygenic photosynthetic organisms and are believed to be essential for the integrity of the membrane system. It remains unclear, however, which part(s) of the galactolipid structure is the key factor for their function, e.g., the sugar moiety and/or the anomeric configuration. To address this issue, several bacterial membrane glycolipid synthase genes have been introduced into cyanobacteria to test for complementation of knocked-out genes involved in galactolipid biosynthesis. In this review, we summarize recent advances in the analyses of sugar species and configurations of glycolipids heterologously synthesized in the thylakoid membrane and discuss their functional importance.
Long interspersed element 1 (L1) retrotransposon sequences are widespread in the human genome, occupying ~500,000 locations. The majority of L1s have lost their retrotransposition capability, although a significant population of human L1s maintains bidirectional transcriptional activity from the internal promoter. While the sense promoter drives transcription of the entire L1 mRNA and leads to L1 retrotransposition, the antisense promoter (ASP) transcribes L1-gene chimeric RNAs that include neighboring exon sequences. Activation mechanisms and functional impacts of L1ASP transcription are thought to vary at every L1ASP location. To explore the locus-specific regulation and function of L1ASP transcription, quantitative methodology is necessary for identifying the genomic positions of highly active L1ASPs on a genome-wide scale. Here, we employed deep-sequencing techniques and built a 3’ RACE-based experimental and bioinformatics protocol, named the L1 antisense transcriptome protocol (LATRAP). In LATRAP, the PCR primer and the read mapping scheme were designed to reduce false positives and negatives, which may have been included as hits in previous cloning studies. LATRAP was here applied to the A549 human lung cancer cell line, and 313 L1ASP loci were detected to have transcriptional activity but differed in the number of mapped reads by four orders of magnitude. This indicates that transcriptional activities of the individual L1ASPs can vary greatly and that only a small population of L1ASP loci is active within individual nuclei. LATRAP is the first experimental method for ranking L1ASPs according to their transcriptional activity and will thus open a new avenue to unveiling the locus-specific biology of L1ASPs.
A full-length cDNA clone encoding grass carp (Ctenopharyngodon idellus) α1-microglobulin/bikunin precursor (Ci-AMBP) was isolated by subtracted differential hybridization screening from a liver cDNA library. The deduced amino acid sequence shared approximately 50% sequence identity with its mammalian counterparts, but more than 90% identity with another fish species. AMBPs are the precursors of the plasma glycoproteins α1-microglobulin (α1m) and bikunin. Both peptide structures and their chromosomal organization were well conserved in Ci-AMBP. The α1m and bikunin polypeptides are separated by the typical tetrapeptide R-A-R-R that provides an endoproteolytic cleavage site for maturation. The genetic organization of domains and functional motifs indicated that Ci-AMBP is a typical member of the lipocalin and Kunitz-type protease inhibitor superfamilies. Expression of the Ci-AMBP gene in different tissues/organs was evaluated using semi-quantitative RT-PCR and, in contrast to the restricted expression in other species, transcripts were detected in a wide range of tissues. The most abundant expression occurred in the secretory organs, which supports the roles of α1m and bikunin in the immune response to diseases and in the stress response.