Autonomous Plasmid Replication inAspergillus nidulans:AMA1 and MATE Elements

Aleksenko, Alexei; Clutterbuck, A. J.

doi:10.1006/fgbi.1997.0980

Cited by 111 publications

(87 citation statements)

References 77 publications

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“…A swoF pyrG strain was transformed with a genomic library constructed in vector pRG3AMA1, which carries the pyr4 gene and the AMA1 sequence for plasmid autonomous replication (1,36). A total of 1,092 pyr prototrophs were screened for growth at restrictive temperature (42°C).…”

Section: Allelesmentioning

confidence: 99%

Aspergillus nidulans swoF Encodes an N -Myristoyl Transferase

Shaw¹,

Momany

Momany³

2002

Eukaryot Cell

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Polar growth is a fundamental process in filamentous fungi and is necessary for disease initiation in many pathogenic systems. Previously, swoF was identified in Aspergillus nidulans as a single-locus, temperaturesensitive (ts) mutant aberrant in both polarity establishment and polarity maintenance. The swoF gene was cloned by complementation of the ts phenotype and sequenced. The derived protein sequence had high identity with N-myristoyl transferases (NMTs) found in fungi, plants, and animals. In addition, wild-type growth at restrictive temperature was partially restored by the addition of myristic acid to the growth medium. Sequencing revealed that the mutation in swoF changes the conserved aspartic acid 369 to a tyrosine. The predicted A. nidulans SwoF protein, SwoFp, was homology modeled based on crystal structures of NMTs from Saccharomyces cerevisiae and Candida albicans. The D369Y swoF mutation is on the opposite face of the protein, distal to the myristoyl coenzyme A and peptide substrate binding sites. In wild-type NMTs, D369 appears to stabilize a structural ␤-strand bend through two hydrogen bonds and an ionic interaction. These stabilizing bonds are abolished in the D369Y mutant. We hypothesize that a substrate of SwoFp must be myristoylated for proper polarity establishment and maintenance. The mutation prevents the proper function of SwoFp at restrictive temperature and thus blocks polar growth.

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Section: Allelesmentioning

confidence: 99%

Aspergillus nidulans swoF Encodes an N -Myristoyl Transferase

Shaw¹,

Momany

Momany³

2002

Eukaryot Cell

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“…AMA1 containing vectors have been designed and can be used for cotransformation with genomic libraries or for the new original instant bank procedure to isolate genes . However, the AMA1 bearing vectors work as replicating plasmids only with some fungi (Aleksenko and Clutterbuck, 1997).…”

Section: Academic Pressmentioning

confidence: 99%

Use of a Linear Plasmid Containing Telomeres as an Efficient Vector for Direct Cloning in The Filamentous FungusPodospora anserina

Barreau

Maya

Turcq

et al. 1998

Fungal Genetics and Biology

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“…It was recovered from a genomic library of A. nidulans during a search for an element that increased transformation efficiency (Gems et al 1991). AMA1-bearing vectors generally have the following properties: (a) a high transformation efficiency that results because these vectors, unlike genomeintegrating vectors, obviate the need for integration into the fungal genome, (b) these vectors localize to nuclei of fungal cells, (c) in Southern blot analysis, these vectors extracted from fungal transformants are detected as bands with electrophoretic mobility similar or identical to circular and/or linear forms of the vector extracted from Escherichia coli, (d) these vectors tend to be lost from fungal lineages under non-selective conditions, and (e) these vectors can be recovered as circular plasmids from E. coli transformed with DNA extract from fungal transformants (Aleksenko and Clutterbuck 1997).…”

Section: Introductionmentioning

confidence: 99%

Transient and multivariate system for transformation of a fungal plant pathogen, Rosellinia necatrix, using autonomously replicating vectors

2012

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Rosellinia necatrix is a fungus that infects a wide range of host plants and ruins a variety of commercially important crops. DNA fragments can be introduced into R. necatrix using conventional protoplast-PEG transformation and genome-integrating vectors; however, transformation efficiency with this strategy is quite low. Therefore, to establish a more effective transformation system for the studies of R. necatrix, an autonomously replicating vector was constructed using AMA1 sequences derived from Aspergillus nidulans, which is distantly related to R. necatrix. Use of this vector with AMA1 sequences increased transformation efficiency in R. necatrix, and the vector was maintained as a plasmid in the transformants. Transient and multivariate functional analyses in R. necatrix were performed using co-transformation of multiple pAMA-H vectors, which each carried either an expression cassette for eGFP, mOrange2, or a geneticin resistance gene. Furthermore, fluorescent proteins expressed from the autonomously replicating vectors were dispersed throughout fungal colonies even though the vectors themselves were restricted to the center of each colony. This intriguing phenomenon indicated that gene products could move from the center to the margin in a colony of the filamentous fungi via a cell-to-cell transport system.

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Autonomous Plasmid Replication inAspergillus nidulans:AMA1 and MATE Elements

Cited by 111 publications

References 77 publications

Aspergillus nidulans swoF Encodes an N -Myristoyl Transferase

Aspergillus nidulans swoF Encodes an N -Myristoyl Transferase

Use of a Linear Plasmid Containing Telomeres as an Efficient Vector for Direct Cloning in The Filamentous FungusPodospora anserina

Transient and multivariate system for transformation of a fungal plant pathogen, Rosellinia necatrix, using autonomously replicating vectors

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