Investigation of the
            <i>malE</i>
            Promoter and MalR, a Positive Regulator of the Maltose Regulon, for an Improved Expression System in Sulfolobus acidocaldarius

Wagner, Michaela; Wagner, Alexander; Ma, Xiaoqing; Kort, Julia Christin; Ghosh, Abhrajyoti; Rauch, Bernadette; Siebers, Bettina; Albers, Sonja‐Verena

doi:10.1128/aem.03050-13

Cited by 32 publications

(38 citation statements)

References 38 publications

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“…We believe that the 15‐fold induction of the amyA promoter under induced conditions (Wagner et al. ), might lead to an overexpression of AglB causing the growth defect by influencing membrane organization.…”

Section: Resultsmentioning

confidence: 97%

AglB, catalyzing the oligosaccharyl transferase step of the archaeal N‐glycosylation process, is essential in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius

Meyer

Albers

2014

MicrobiologyOpen

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Sulfolobus acidocaldarius, a thermo-acidophilic crenarchaeon which grows optimally at 76°C and pH 3, exhibits an astonishing high number of N-glycans linked to the surface (S-) layer proteins. The S-layer proteins as well as other surface-exposed proteins are modified via N-glycosylation, in which the oligosaccharyl transferase AglB catalyzes the final step of the transfer of the glycan tree to the nascent protein. In this study, we demonstrated that AglB is essential for the viability of S. acidocaldarius. Different deletion approaches, that is, markerless in-frame deletion as well as a marker insertion were unsuccessful to create an aglB deletion mutant. Only the integration of a second aglB gene copy allowed the successful deletion of the original aglB.

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Section: Resultsmentioning

confidence: 97%

AglB, catalyzing the oligosaccharyl transferase step of the archaeal N‐glycosylation process, is essential in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius

Meyer

Albers

2014

MicrobiologyOpen

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“…To this end, both genes were cloned into the expression vectors pSVAmZ-SH10 and pSVA1551, respectively, under the control of the inducible maltose (mal) promotor from S acidocaldarius 38. After growth of the transformants in liquid culture in the presence of dextrin as an inducer, in vivo labelling at 78 °C and pH 3.0 with either FP≡ or NP≡ with or without preincubation with 50 μM paraoxon led to the re-appearance of the 38 kDa band (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…38). To this end and to reduce the size of the pRN1 backbone, five backbone fragments with overlapping ends were amplified via PCR using the ‘mod_ Bam HI del' primers.…”

Section: Methodsmentioning

confidence: 99%

Activity-based protein profiling as a robust method for enzyme identification and screening in extremophilic Archaea

et al. 2017

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Archaea are characterized by a unique life style in often environmental extremes but their thorough investigation is currently hampered by a limited set of suitable in vivo research methodologies. Here, we demonstrate that in vivo activity-based protein profiling (ABPP) may be used to sensitively detect either native or heterogeneously expressed active enzymes in living archaea even under these extreme conditions. In combination with the development of a genetically engineered archaeal screening strain, ABPP can furthermore be used in functional enzyme screenings from (meta)genome samples. We anticipate that our ABPP approach may therefore find application in basic archaeal research but also in the discovery of novel enzymes from (meta)genome libraries.

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“…For the quantification of the β‐galactosidase activity the following equation was used (Miller, ; Wagner et al ., ):

normalM normali normall normall normale normalr = \frac{60 000 * (|, A 410 (|, t 2 - t 1) - autolysis A 410 (|, t 2 - t 1)) * 7}{normalt normali normalm normale (|, normals) * normalv normalo normall normalu normalm normale normalo normalf normalt normalh normale normals normala normalm normalp normall normale (|, normalm normall) * normalp normalr normalo normalt normale normali normaln normalc normalo normaln normalc normale normaln normalt normalr normala normalt normali normalo normaln true(normalm normalg / normalm normall true)}

…”

Section: Methodsmentioning

confidence: 99%

Plasticity of archaeal C/D box sRNA biogenesis

Tripp

Martin

Orell

et al. 2016

Molecular Microbiology

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SummaryArchaeal and eukaryotic organisms contain sets of C/D box s(no)RNAs with guide sequences that determine ribose 2 0 -O-methylation sites of target RNAs. The composition of these C/D box sRNA sets is highly variable between organisms and results in varying RNA modification patterns which are important for ribosomal RNA folding and stability. Little is known about the genomic organization of C/D box sRNA genes in archaea. Here, we aimed to obtain first insights into the biogenesis of these archaeal C/D box sRNAs and analyzed the genetic context of more than 300 archaeal sRNA genes. We found that the majority of these genes do not possess independent promoters but are rather located at positions that allow for co-transcription with neighboring genes and their start or stop codons were frequently incorporated into the conserved boxC and D motifs. The biogenesis of plasmidencoded C/D box sRNA variants was analyzed in vivo in Sulfolobus acidocaldarius. It was found that C/D box sRNA maturation occurs independent of their genetic context and relies solely on the presence of intact RNA kink-turn structures. The observed plasticity of C/D box sRNA biogenesis is suggested to enable their accelerated evolution and, consequently, allow for adjustments of the RNA modification landscape.

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Investigation of the malE Promoter and MalR, a Positive Regulator of the Maltose Regulon, for an Improved Expression System in Sulfolobus acidocaldarius

Cited by 32 publications

References 38 publications

AglB, catalyzing the oligosaccharyl transferase step of the archaeal N‐glycosylation process, is essential in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius

AglB, catalyzing the oligosaccharyl transferase step of the archaeal N‐glycosylation process, is essential in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius

Activity-based protein profiling as a robust method for enzyme identification and screening in extremophilic Archaea

Plasticity of archaeal C/D box sRNA biogenesis

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