Propionyl coenzyme A carboxylase is required for development of Myxococcus xanthus

Kimura, Yoshinobu; Sato, Rikishi; Mimura, Kayoko; Sato, Masayuki

doi:10.1128/jb.179.22.7098-7102.1997

Cited by 9 publications

(12 citation statements)

References 41 publications

(34 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These four genes are predicted to encode homologs of the Pseudomonas aeruginosa genes fimT, pilW, pilV, and pilY1, which contribute to the stability of type IV pili Mattick 1995, 1997;Alm et al 1996;Carbonnelle et al 2005). The second of these is in a gene, MXAN3759, predicted to encode one of two propionyl-CoA carboxylases made by M. xanthus (Kimura et al 1997). The third of these is in a gene, MXAN3797, predicted to encode a short-chain acyl-CoA dehydrogenase.…”

Section: Resultsmentioning

confidence: 99%

“…A defect in its paralogue, pccB (MXAN1113), results in a reduction in the relative amount of long-chain fatty acids present in the membranes of vegetative and developing cells, but not in their abolition. Thus, although a pccB mutant can aggregate, it cannot form heat-resistant spores in response to carbon starvation (Kimura et al 1997), a phenotype that can be masked partially by the addition of methylmalonyl-CoA to developing cells. The fact that a second propionyl-CoA acyl carboxylase isozyme also is required for social motility underscores the importance of long-chain fatty acids in the integrity of the M. xanthus membrane.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Transposon Insertions ofmagellan-4That Impair Social Gliding Motility inMyxococcus xanthus

Youderian

Hartzell

2006

Genetics

View full text Add to dashboard Cite

Myxococcus xanthus has two different mechanisms of motility, adventurous (A) motility, which permits individual cells to glide over solid surfaces, and social (S) motility, which permits groups of cells to glide. To identify the genes involved in S-gliding motility, we mutagenized a DaglU (A ÿ ) strain with the defective transposon, magellan-4, and screened for S ÿ mutants that form nonmotile colonies. Sequence analysis of the sites of the magellan-4 insertions in these mutants and the alignment of these sites with the M. xanthus genome sequence show that two-thirds of these insertions lie within 27 of the 37 nonessential genes known to be required for social motility, including those necessary for the biogenesis of type IV pili, exopolysaccharide, and lipopolysaccharide. The remaining insertions also identify 31 new, nonessential genes predicted to encode both structural and regulatory determinants of S motility. These include three tetratricopeptide repeat proteins, several regulators of transcription that may control the expression of genes involved in pilus extension and retraction, and additional enzymes involved in polysaccharide metabolism. Three insertions that abolish S motility lie within genes predicted to encode glycolytic enzymes, suggesting that the signal for pilus retraction may be a simple product of exopolysaccharide catabolism.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Transposon Insertions ofmagellan-4That Impair Social Gliding Motility inMyxococcus xanthus

Youderian

Hartzell

2006

Genetics

View full text Add to dashboard Cite

show abstract

“…Thus, evidence for the production of methylmalonyl-CoA cannot be deduced from the polyketides that are made naturally from this host. Work studying the developmental life cycle of M. xanthus has revealed the presence of the genes for propionyl-CoA carboxylase (16,17), an enzyme used to synthesize methyl malonyl-CoA. Thus, M. xan-…”

mentioning

confidence: 99%

Heterologous Expression of Epothilone Biosynthetic Genes in Myxococcus xanthus

Julien¹,

Shah²

2002

Antimicrob Agents Chemother

129

View full text Add to dashboard Cite

Epothilones are potential anticancer drugs that stabilize microtubules in a manner similar to paclitaxel (Taxol). Epothilones are produced from the myxobacterium Sorangium cellulosum, which has a 16-h doubling time and produces only milligram-per-liter amounts of epothilone A and epothilone B. Furthermore, genetic manipulation of S. cellulosum is difficult. To produce epothilones in a more genetically amenable and rapidly growing host, we chose the closely related and best-characterized myxobacteria Myxococcus xanthus. We inserted 65.4 kb of S. cellulosum DNA that encompassed the entire epothilone gene cluster into the chromosome of M. xanthus by a series of homologous recombination events. The resulting strain produced epothilones A and B. Construction of a strain that contained a mutation in epoK, the P450 epoxidase, resulted in production of epothilones C and D.Epothilones are potent cytotoxic macrocyclic lactones that show promise as anticancer drugs (21,22,26). The mechanism of action is similar to the anticancer drug paclitaxel (Taxol); both bind and stabilize microtubules, which leads to cell death (3). Epothilones have superior features relative to paclitaxel. First, epothilones are more water soluble. This may enable a formulation without the use of the solubilizing agent cremophor, currently used in formulations of paclitaxel. Cremophor, on its own, can affect cardiac function and cause severe hypersensitivity (25). Second, epothilones are effective against tumors resistant to paclitaxel (3, 31). These advantages make epothilones likely successors to paclitaxel.The need for sufficient material is a major obstacle to the development of epothilones as marketable drugs. The total synthesis of epothilone A and epothilone B has been accomplished (2, 32). However, the number of steps required for synthesis of these molecules precludes this as an economical method of production. Fermentation methods thus remain the favored route of production.Epothilones are produced from the gram-negative myxobacterium Sorangium cellulosum (8). The reported yields of epothilones from S. cellulosum strain So ce90 are approximately 20 mg of epothilone A per liter and 10 mg of epothilone B per liter (8). A disadvantage of S. cellulosum is the relatively long doubling time, which is approximately 16 h and is the longest of all myxobacteria. Furthermore, S. cellulosum is difficult to engineer, due to the low efficiency of introducing DNA into the bacterium (11) and the limited number of molecular tools and markers that have been developed.Recently, the epothilone biosynthetic gene cluster was sequenced, and the genes were introduced into Streptomyces coelicolor, a common host used for production of a variety of polyketides from actinomycetes (33). The heterologous strain produced small quantities of epothilones; production of epothilones in S. coelicolor may have a cytotoxic effect (L. Tang et al., unpublished data). Thus, an alternative heterologous host is desired.We speculated that a superior expression host for epothilones mi...

show abstract

“…AccB showed high degrees of sequence similarities with the ␣ subunit of Veillonella parvula methylmalonyl-CoA decarboxylase (27% identity) (18), the ␤ subunit of M. xanthus propionyl-CoA carboxylase (28% identity) (20), and the carboxyltransferase ␣ and ␤ subunits of E. coli acetyl-CoA carboxylase (19 and 13% identity, respectively) (25) (Fig. 3).…”

Section: Resultsmentioning

confidence: 99%

“…letion mutant was unable to sporulate under conditions of nutrient starvation (20). The developing cells of the mutant also showed reduced levels of long-chain fatty acids compared to wild-type cells.…”

mentioning

confidence: 99%

Molecular Cloning and Characterization of Two Genes for the Biotin Carboxylase and Carboxyltransferase Subunits of Acetyl Coenzyme A Carboxylase in Myxococcus xanthus

et al. 2000

View full text Add to dashboard Cite

We have cloned a DNA fragment from a genomic library of Myxococcus xanthus using an oligonucleotide probe representing conserved regions of biotin carboxylase subunits of acetyl coenzyme A (acetyl-CoA) carboxylases. The fragment contained two open reading frames (ORF1 and ORF2), designated the accB and accA genes, capable of encoding a 538-amino-acid protein of 58.1 kDa and a 573-amino-acid protein of 61.5 kDa, respectively. The protein (AccA) encoded by the accA gene was strikingly similar to biotin carboxylase subunits of acetyl-CoA and propionyl-CoA carboxylases and of pyruvate carboxylase. The putative motifs for ATP binding, CO 2 fixation, and biotin binding were found in AccA. The accB gene was located upstream of the accA gene, and they formed a two-gene operon. The protein (AccB) encoded by the accB gene showed high degrees of sequence similarity with carboxyltransferase subunits of acetyl-CoA and propionyl-CoA carboxylases and of methylmalonyl-CoA decarboxylase. Carboxybiotin-binding and acyl-CoA-binding domains, which are conserved in several carboxyltransferase subunits of acyl-CoA carboxylases, were found in AccB. An accA disruption mutant showed a reduced growth rate and reduced acetyl-CoA carboxylase activity compared with the wild-type strain. Western blot analysis indicated that the product of the accA gene was a biotinylated protein that was expressed during the exponential growth phase. Based on these results, we propose that this M. xanthus acetyl-CoA carboxylase consists of two subunits, which are encoded by the accB and accA genes, and occupies a position between prokaryotic and eukaryotic acetyl-CoA carboxylases in terms of evolution.

show abstract

Propionyl coenzyme A carboxylase is required for development of Myxococcus xanthus

Cited by 9 publications

References 41 publications

Transposon Insertions ofmagellan-4That Impair Social Gliding Motility inMyxococcus xanthus

Transposon Insertions ofmagellan-4That Impair Social Gliding Motility inMyxococcus xanthus

Heterologous Expression of Epothilone Biosynthetic Genes in Myxococcus xanthus

Molecular Cloning and Characterization of Two Genes for the Biotin Carboxylase and Carboxyltransferase Subunits of Acetyl Coenzyme A Carboxylase in Myxococcus xanthus

Contact Info

Product

Resources

About