scholarly journals glkA Is Involved in Glucose Repression of Chitinase Production in Streptomyces lividans

1998 ◽  
Vol 180 (11) ◽  
pp. 2911-2914 ◽  
Author(s):  
Akihiro Saito ◽  
Takeshi Fujii ◽  
Tadakatsu Yoneyama ◽  
Kiyotaka Miyashita
Keyword(s):  
Sole Carbon Source ◽  
Glucose Repression ◽  
Streptomyces Coelicolor ◽  
Streptomyces Lividans ◽  
Transcriptional Analysis ◽  
Copy Number Plasmid ◽  
Low Copy Number ◽  
Chitinase Production ◽  
Dna Fragment ◽  
Glucose Kinase

ABSTRACT Chitinase production in Streptomyces lividans is induced by chitin and repressed in the presence of glucose. A mutant ofS. lividans TK24, strain G015, which was defective in glucose repression of chitinase production, was obtained by screening colonies for zones of clearing on colloidal chitin agar plates containing 1.0% (wt/vol) glucose. The transcriptional analysis ofchiA in G015 with xylE, which encodes catechol 2,3-dioxygenase, as a reporter gene showed that the transcription from the chiA promoter of S. lividans TK24 occurred regardless of the presence of glucose. G015 was resistant to 2-deoxyglucose (2-DOG) and did not utilize glucose as a sole carbon source. When a DNA fragment containing glkA, a gene for glucose kinase, of Streptomyces coelicolor A3(2) was introduced into strain G015 on a low-copy-number plasmid, the sensitivity to 2-DOG, the ability to utilize glucose, and the glucose repression of chitinase production were restored. These results indicate that glkA is involved in glucose repression of chitinase production in S. lividans TK24.

Glucose repression in Streptomyces coelicolor A3(2): a likely regulatory role for glucose kinase

1994 ◽  
Vol 244 (2) ◽  
pp. 135-143 ◽  
Author(s):  
Susan Angell ◽  
Cinzia G. Lewis ◽  
Mark J. Buttner ◽  
Mervyn J. Bibb
Keyword(s):  
Glucose Repression ◽  
Streptomyces Coelicolor ◽  
Regulatory Role ◽  
Glucose Kinase

Possible involvement of the sco2127 gene product in glucose repression of actinorhodin production in Streptomyces coelicolor

2012 ◽  
Vol 58 (10) ◽  
pp. 1195-1201 ◽  
Author(s):  
Angela Forero ◽  
Mauricio Sánchez ◽  
Adán Chávez ◽  
Beatriz Ruiz ◽  
Romina Rodríguez-Sanoja ◽  
...  
Keyword(s):  
Glucose Repression ◽  
Antibiotic Production ◽  
Streptomyces Coelicolor ◽  
Negative Regulator ◽  
Wild Type ◽  
Wild Type Phenotype ◽  
Glucose Sensitivity ◽  
Mutant Complementation ◽  
Glucose Kinase ◽  
Actinorhodin Production

Streptomyces coelicolor mutants resistant to 2-deoxyglucose are insensitive to carbon catabolite repression (CCR). Total reversion to CCR sensitivity is observed by mutant complementation with a DNA region harboring both glucose kinase glkA gene and the sco2127 gene. The sco2127 is located upstream of glkA and encodes a putative protein of 20.1 kDa. In S. coelicolor, actinorhodin production is subject to glucose repression. To explore the possible involvement of both SCO2127 and glucose kinase (Glk) in the glucose sensitivity of actinorhodin production, this effect was evaluated in a wild-type S. coelicolor A3(2) M145 strain and a sco2127 null mutant (Δsco2127) derived from this wild-type strain. In comparison with strain M145, actinorhodin production by the mutant was insensitive to glucose repression. Under repressive conditions, only minor differences were observed in glucose utilization and Glk production between these strains. SCO2127 was detected mainly during the first 36 h of fermentation, just before the onset of antibiotic production, and its synthesis was not related to a particular carbon source. The glucose sensitivity of antibiotic production was restored to wild-type phenotype by transformation with an integrative plasmid containing sco2127. Our results support the hypothesis that SCO2127 is a negative regulator of actinorhodin production and suggest that the effect is independent of Glk.

scholarly journals Modulation of Actinorhodin Biosynthesis in Streptomyces lividans by Glucose Repression of afsR2 Gene Transcription

2001 ◽  
Vol 183 (7) ◽  
pp. 2198-2203 ◽  
Author(s):  
Eung-Soo Kim ◽  
Hee-Jeon Hong ◽  
Cha-Yong Choi ◽  
Stanley N. Cohen
Keyword(s):  
Carbon Source ◽  
Glucose Repression ◽  
Streptomyces Coelicolor ◽  
Bacterial Species ◽  
Morphological Differentiation ◽  
Aerial Mycelium ◽  
Biosynthetic Gene Cluster ◽  
Streptomyces Lividans ◽  
Deep Blue ◽  
Solid Media

ABSTRACT While the biosynthetic gene cluster encoding the pigmented antibiotic actinorhodin (ACT) is present in the two closely related bacterial species, Streptomyces lividans andStreptomyces coelicolor, it normally is expressed only inS. coelicolor—generating the deep-blue colonies responsible for the S. coelicolor name. However, multiple copies of the two regulatory genes, afsR andafsR2, activate ACT production in S. lividans, indicating that this streptomycete encodes a functional ACT biosynthetic pathway. Here we report that the occurrence of ACT biosynthesis in S. lividans is determined conditionally by the carbon source used for culture. We found that the growth ofS. lividans on solid media containing glucose prevents ACT production in this species by repressing the synthesis ofafsR2 mRNA; a shift to glycerol as the sole carbon source dramatically relieved this repression, leading to extensive ACT synthesis and obliterating this phenotypic distinction between S. lividans and S. coelicolor. Transcription from theafsR2 promoter during growth in glycerol was dependent onafsR gene function and was developmentally regulated, occurring specifically at the time of aerial mycelium formation and coinciding temporally with the onset of ACT production. In liquid media, where morphological differentiation does not occur, ACT production in the absence of glucose increased as S. lividans cells entered stationary phase, but unlike ACT biosynthesis on solid media, occurred by a mechanism that did not require either afsR or afsR2. Our results identify parallel medium-dependent pathways that regulate ACT biosynthesis in S. lividans and further demonstrate that the production of this antibiotic in S. lividans grown on agar can be modulated by carbon source through the regulation ofafsR2 mRNA synthesis.

scholarly journals An Additional Regulatory Gene for Actinorhodin Production in Streptomyces lividans Involves a LysR-Type Transcriptional Regulator

1999 ◽  
Vol 181 (14) ◽  
pp. 4353-4364 ◽  
Author(s):  
Oscar H. Martínez-Costa ◽  
Angel J. Martín-Triana ◽  
Eduardo Martínez ◽  
Miguel A. Fernández-Moreno ◽  
Francisco Malpartida
Keyword(s):  
Streptomyces Coelicolor ◽  
Regulatory Gene ◽  
Streptomyces Lividans ◽  
Amino Acid Sequences ◽  
Open Reading Frames ◽  
Transcriptional Analysis ◽  
The Family ◽  
The Right ◽  
Different Sources ◽  
Transcription Expression

The sequence of a 4.8-kbp DNA fragment adjacent to the right-hand end of the actinorhodin biosynthetic (act) cluster downstream of actVB-orf6 from Streptomyces coelicolor A3(2) reveals six complete open reading frames, namedorf7 to orf12. The deduced amino acid sequences from orf7, orf10, and orf11 show significant similarities with the following products in the databases: a putative protein from the S. coelicolor SCP3 plasmid, LysR-type transcriptional regulators, and proteins belonging to the family of short-chain dehydrogenases/reductases, respectively. The deduced product of orf8 reveals low similarities with several methyltransferases from different sources, whileorf9 and orf12 products show no similarities with other known proteins. Disruptions of orf10 andorf11 genes in S. coelicolor appear to have no significant effect on the production of actinorhodin. Nevertheless, disruption or deletion of orf10 in Streptomyces lividans causes actinorhodin overproduction. The introduction of extra copies of orf10 and orf11 genes in anS. coelicolor actIII mutant restores the ability to produce actinorhodin. Transcriptional analysis and DNA footprinting indicate that Orf10 represses its own transcription and regulatesorf11 transcription, expression of which might require the presence of an unknown inducer. No DNA target for Orf10 protein was found within the act cluster.

scholarly journals In Vivo Analysis of HPr Reveals a Fructose-Specific Phosphotransferase System That Confers High-Affinity Uptake in Streptomyces coelicolor

2003 ◽  
Vol 185 (3) ◽  
pp. 929-937 ◽  
Author(s):  
Harald Nothaft ◽  
Stephan Parche ◽  
Annette Kamionka ◽  
Fritz Titgemeyer
Keyword(s):  
Carbon Source ◽  
Glucose Repression ◽  
Streptomyces Coelicolor ◽  
Carbon Sources ◽  
Glycerol Kinase ◽  
Transcriptional Analysis ◽  
Phosphotransferase System ◽  
Gram Positive ◽  
High Affinity

ABSTRACT HPr, the histidine-containing phosphocarrier protein of the bacterial phosphotransferase system (PTS), serves multiple functions in carbohydrate uptake and carbon source regulation in low-G+C-content gram-positive bacteria and in gram-negative bacteria. To assess the role of HPr in the high-G+C-content gram-positive organism Streptomyces coelicolor, the encoding gene, ptsH, was deleted. The ptsH mutant BAP1 was impaired in fructose utilization, while growth on other carbon sources was not affected. Uptake assays revealed that BAP1 could not transport appreciable amounts of fructose, while the wild type showed inducible high-affinity fructose transport with an apparent Km of 2 μM. Complementation and reconstitution experiments demonstrated that HPr is indispensable for a fructose-specific PTS activity. Investigation of the putative fruKA gene locus led to identification of the fructose-specific enzyme II permease encoded by the fruA gene. Synthesis of HPr was not specifically enhanced in fructose-grown cells and occurred also in the presence of non-PTS carbon sources. Transcriptional analysis of ptsH revealed two promoters that are carbon source regulated. In contrast to what happens in other bacteria, glucose repression of glycerol kinase was still operative in a ptsH background, which suggests that HPr is not involved in general carbon regulation. However, fructose repression of glycerol kinase was lost in BAP1, indicating that the fructose-PTS is required for transduction of the signal. This study provides the first molecular genetic evidence of a physiological role of the PTS in S. coelicolor.

scholarly journals Backbone assignments, and effect of Asn deamidation, of the N-terminal region of the partitioning protein IncC1 from the plasmid RK2

2021 ◽  
Author(s):  
M. Fayyaz Rehman ◽  
M. Jeeves ◽  
E. I. Hyde
Keyword(s):  
Amino Acids ◽  
Chemical Shift Assignments ◽  
Backbone Assignments ◽  
Intrinsically Disordered ◽  
Nmr Chemical Shift Assignments ◽  
Protein Partner ◽  
Copy Number Plasmid ◽  
Low Copy Number ◽  
Plasmid Rk2

AbstractIncC from the low-copy number plasmid RK2, is a member of the ParA family of proteins required for partitioning DNA in many bacteria and plasmids. It is an ATPase that binds DNA and its ParB protein partner, KorB. Together, the proteins move replicated DNA to appropriate cellular positions, so that each daughter cell inherits a copy on cell division. IncC from RK2 is expressed in two forms. IncC2 is homologous to bacterial ParA proteins, while IncC1 has an N-terminal extension of 105 amino acids and is similar in length to ParA homologues in other plasmids. We have been examining the role of this extension, here called IncC NTD. We present its backbone NMR chemical shift assignments and show that it is entirely intrinsically disordered. The assignments were achieved using C-detected, CON-based spectra, complemented by HNN spectra to obtain connectivities from three adjacent amino acids. We also observed evidence of deamidation of the protein at a GNGG sequence, to give isoAsp, giving 2 sets of peaks for residues up to 5 amino acids on either side of the modification. We have assigned resonances from around the position of modification for this form of the protein.

scholarly journals Regulation of Sporangium Formation by BldD in the Rare Actinomycete Actinoplanes missouriensis

2017 ◽  
Vol 199 (12) ◽  
Author(s):  
Yoshihiro Mouri ◽  
Kenji Konishi ◽  
Azusa Fujita ◽  
Takeaki Tezuka ◽  
Yasuo Ohnishi
Keyword(s):  
Vegetative Growth ◽  
Streptomyces Coelicolor ◽  
Morphological Differentiation ◽  
Transcriptional Regulator ◽  
Saccharopolyspora Erythraea ◽  
Transcriptional Analysis ◽  
Morphological Development ◽  
Sequencing Analysis ◽  
Content Type ◽  
Biological Studies

ABSTRACT The rare actinomycete Actinoplanes missouriensis forms sporangia, including hundreds of flagellated spores that start swimming as zoospores after their release. Under conditions suitable for vegetative growth, zoospores stop swimming and germinate. A comparative proteome analysis between zoospores and germinating cells identified 15 proteins that were produced in larger amounts in germinating cells. They include an orthologue of BldD (herein named AmBldD [BldD of A. missouriensis]), which is a transcriptional regulator involved in morphological development and secondary metabolism in Streptomyces. AmBldD was detected in mycelia during vegetative growth but was barely detected in mycelia during the sporangium-forming phase, in spite of the constant transcription of AmbldD throughout growth. An AmbldD mutant started to form sporangia much earlier than the wild-type strain, and the resulting sporangia were morphologically abnormal. Recombinant AmBldD bound a palindromic sequence, the AmBldD box, located upstream from AmbldD. 3′,5′-Cyclic di-GMP significantly enhanced the in vitro DNA-binding ability of AmBldD. A chromatin immunoprecipitation-sequencing analysis and an in silico search for AmBldD boxes revealed that AmBldD bound 346 genomic loci that contained the 19-bp inverted repeat 5′-NN(G/A)TNACN(C/G)N(G/C)NGTNA(C/T)NN-3′ as the consensus AmBldD-binding sequence. The transcriptional analysis of 27 selected AmBldD target gene candidates indicated that AmBldD should repress 12 of the 27 genes, including bldM, ssgB, whiD, ddbA, and wblA orthologues. These genes are involved in morphological development in Streptomyces coelicolor A3(2). Thus, AmBldD is a global transcriptional regulator that seems to repress the transcription of tens of genes during vegetative growth, some of which are likely to be required for sporangium formation. IMPORTANCE The rare actinomycete Actinoplanes missouriensis undergoes complex morphological differentiation, including sporangium formation. However, almost no molecular biological studies have been conducted on this bacterium. BldD is a key global regulator involved in the morphological development of streptomycetes. BldD orthologues are highly conserved among sporulating actinomycetes, but no BldD orthologues, except one in Saccharopolyspora erythraea, have been studied outside the streptomycetes. Here, it was revealed that the BldD orthologue AmBldD is essential for normal developmental processes in A. missouriensis. The AmBldD regulon seems to be different from the BldD regulon in Streptomyces coelicolor A3(2), but they share four genes that are involved in morphological differentiation in S. coelicolor A3(2).

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