Activation of the Promoter of the Fengycin Synthetase Operon by the UP Element

Ke, Wan-Ju; Chang, Ban‐Yang; Lin, Tsuey-Pin; Liu, Shih‐Tung

doi:10.1128/jb.00255-09

Cited by 14 publications

(8 citation statements)

References 52 publications

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“…In this way, binding of the specific ligand to the RNA riboswitch may result in structural changes that could turn a gene on or off ( Coppins et al ., 2007 ). Furthermore, some UP elements, DNA fragments located upstream from the tss and usually with a high A/T content, have also been involved in transcriptional regulation in Gram‐positive bacteria ( Estrem et al ., 1998 ; Ke et al ., 2009 ). To determine the putative role of the large inverted repeats on Bbr_0838 expression, gusA fusions lacking DNA regions of variable length immediately upstream from Bbr_0838, but retaining the region encompassing the presumed −35 and −10 sequences and the transcriptional start site, were constructed and their bile‐inducibility measured through GusA activity ( Fig.…”

Section: Resultsmentioning

confidence: 99%

A bile‐inducible membrane protein mediates bifidobacterial bile resistance

Ruíz

Motherway

Zomer

et al. 2012

Microbial Biotechnology

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SummaryBbr_0838 from Bifidobacterium breve UCC2003 is predicted to encode a 683 residue membrane protein, containing both a permease domain that displays similarity to transporters belonging to the major facilitator superfamily, as well as a CBS (cystathionine beta synthase) domain. The high level of similarity to bile efflux pumps from other bifidobacteria suggests a significant and general role for Bbr_0838 in bile tolerance. Bbr_0838 transcription was shown to be monocistronic and strongly induced upon exposure to bile. Further analysis delineated the transcriptional start site and the minimal region required for promoter activity and bile regulation. Insertional inactivation of Bbr_0838 in B. breve UCC2003 resulted in a strain, UCC2003:838800, which exhibited reduced survival upon cholate exposure as compared with the parent strain, a phenotype that was reversed when a functional, plasmid‐encoded Bbr_0838 gene was introduced into UCC2003:838800. Transcriptome analysis of UCC2003:838800 grown in the presence or absence of bile demonstrated that transcription of Bbr_0832, which is predicted to encode a macrolide efflux transporter gene, was significantly increased in the presence of bile, representing a likely compensatory mechanism for bile removal in the absence of Bbr_0838. This study represents the first in‐depth analysis of a bile‐inducible locus in bifidobacteria, identifying a key gene relevant for bifidobacterial bile tolerance.

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Section: Resultsmentioning

confidence: 99%

A bile‐inducible membrane protein mediates bifidobacterial bile resistance

Ruíz

Motherway

Zomer

et al. 2012

Microbial Biotechnology

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“…The promoters that are regulated by an UP element include E. coli rrnB P1 and that of the Bacillus subtilis fengycin synthetase operon (24,36). In fact, an UP element contains two subsites where the two ␣CTDs in RNA polymerase bind (17,20).…”

Section: Discussionmentioning

confidence: 99%

“…Probe p-msps had the same sequence as p-sps except that the region between nt 380 and 388 was changed from 5Ј-AAGATCTTC to 5Ј-CCACGAGGA. ␣CTD (250 ng), purified from E. coli BL21(pWCTD) using nickel-nitrilotriacetic acid (Ni-NTA) agarose beads (Qiagen) according to a method described elsewhere (24), was mixed with 1 g of a biotinylated probe. The beads were washed and captured using streptavidin MagneSphere paramagnetic particles (Promega).…”

Section: Vol 192 2010 Activation Of Rnaii Promoter 3655mentioning

confidence: 99%

A Sequence That Affects the Copy Number and Stability of pSW200 and ColE1

Liu

2010

J Bacteriol

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Pantoea stewartii SW2 contains 13 plasmids. One of these plasmids, pSW200, has a replicon that resembles that of ColE1. This study demonstrates that pSW200 contains a 9-bp UP element, 5-AAGATCTTC, which is located immediately upstream of the ؊35 box in the RNAII promoter. A transcriptional fusion study reveals that substituting this 9-bp sequence reduces the activity of the RNAII promoter by 78%. The same mutation also reduced the number of plasmid copies from 13 to 5, as well as the plasmid stability. When a similar sequence in a ColE1 derivative, pYCW301, is mutated, the copy number of the plasmid also declines from 34 to 16 per cell. Additionally, inserting this 9-bp sequence stabilizes an unstable pSW100 derivative, pSW142K, which also contains a replicon resembling that of ColE1, indicating the importance of this sequence in maintaining the stability of the plasmid. In conclusion, the 9-bp sequence upstream of the ؊35 box in the RNAII promoter is required for the efficient synthesis of RNAII and maintenance of the stability of the plasmids in the ColE1 family.

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“…The proteins were separated by in a 6% SDS-polyacrylamide gel and analyzed by immunoblotting using anti-RNA polymerase β subunit antibody and anti-σ 70 antibody (Abcam, Cambridge, UK) [37]. A biotin-labeled probe, 167, which contained a non-promoter sequence from the fen operon [47] was used as a negative control.…”

Section: Methodsmentioning

confidence: 99%

A Repeat Sequence Causes Competition of ColE1-Type Plasmids

Lin

Liu

2013

PLoS ONE

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Plasmid pSW200 from Pantoea stewartii contains 41 copies of 15-bp repeats and has a replicon that is homologous to that of ColE1. Although deleting the repeats (pSW207) does not change the copy number and stability of the plasmid. The plasmid becomes unstable and is rapidly lost from the host when a homoplasmid with the repeats (pSW201) is present. Deleting the repeats is found to reduce the transcriptional activity of RNAIp and RNAIIp by about 30%, indicating that the repeats promote the transcription of RNAI and RNAII, and how the RNAI that is synthesized by pSW201 inhibits the replication of pSW207. The immunoblot analysis herein demonstrates that RNA polymerase β subunit and σ70 in the lysate from Escherichia coli MG1655 bind to a biotin-labeled DNA probe that contains the entire sequence of the repeat region. Electrophoretic mobility shift assay also reveals that purified RNA polymerase shifts a DNA probe that contains four copies of the repeats. These results thus obtained reveal that RNA polymerase holoenzyme binds to the repeats. The repeats also exchange RNA polymerase with RNAIp and RNAIIp in vitro, revealing the mechanism by which the transcription is promoted. This investigation elucidates a mechanism by which a plasmid prevents the invasion of an incompatible plasmid and maintains its stability in the host cell during evolution.

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Activation of the Promoter of the Fengycin Synthetase Operon by the UP Element

Cited by 14 publications

References 52 publications

A bile‐inducible membrane protein mediates bifidobacterial bile resistance

A bile‐inducible membrane protein mediates bifidobacterial bile resistance

A Sequence That Affects the Copy Number and Stability of pSW200 and ColE1

A Repeat Sequence Causes Competition of ColE1-Type Plasmids

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