The isolation and characterization of the pyruvate kinase-encoding gene from the yeast Yarrowia lipolytica

Strick, Christine A.; James, Larry C.; O'Donnell, M.M.; Gollaher, M.G.; Franke, Arthur E.

doi:10.1016/0378-1119(92)90249-o

Cited by 19 publications

(12 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Sequencing of this fragment revealed an open reading frame of 2,577 bp interrupted by one intron, which is found between codons 3 and 4 (nucleotides ϩ10 to ϩ666 from the A residue of the potential initiating codon). The putative 5Ј splice donor (GTGAGTPu) and 3Ј splice acceptor (TACTAACNCAG) sequences are identical to the motifs found in other Y. lipolytica genes (33,56,58) (Fig. 2), and analysis of YlBEM1 cDNA showed that these sequences are functional (data not shown).…”

Section: Resultsmentioning

confidence: 58%

Isolation and Characterization of YlBEM1 , a Gene Required for Cell Polarization and Differentiation in the Dimorphic Yeast Yarrowia lipolytica

Hurtado

Rachubinski

2002

Eukaryot Cell

View full text Add to dashboard Cite

The ability to switch between a unicellular yeast form and different filamentous forms (fungal dimorphism) is an important attribute of most pathogenic fungi. Dimorphism involves a series of events that ultimately result in dramatic changes in the polarity of cell growth in response to environmental factors. We have isolated and characterized YlBEM1, a gene encoding a protein of 639 amino acids that is essential for the yeast-to-hypha transition in the yeast Yarrowia lipolytica and whose transcription is significantly increased during this event. Cells with deletions of YlBEM1 are viable but show substantial alterations in morphology, disorganization of the actin cytoskeleton, delocalization of cortical actin and chitin deposition, multinucleation, and loss of mating ability, thus pointing to a major role for YlBEM1 in the regulation of cell polarity and morphogenesis in this fungus. This role is further supported by the localization of YlBem1p, which, like cortical actin, appears to be particularly abundant at sites of growth of yeast, hyphal, and pseudohyphal cells. In addition, the potential involvement of YlBem1p in septum formation and/or cytokinesis is suggested by the concentration of a green fluorescent protein-tagged version of this protein at the mother-bud neck during the last stages of cell division. Interestingly, overexpression of MHY1, YlRAC1, or YlSEC31, three genes involved in filamentous growth of Y. lipolytica, induced hyphal growth of bem1 null mutant cells.

show abstract

Section: Resultsmentioning

confidence: 58%

Isolation and Characterization of YlBEM1 , a Gene Required for Cell Polarization and Differentiation in the Dimorphic Yeast Yarrowia lipolytica

Hurtado

Rachubinski

2002

Eukaryot Cell

View full text Add to dashboard Cite

show abstract

“…2). The putative 5Ј-splice donor sequences of both introns (GTA AGTPu) diverge at the third and fourth positions from the GTGAGTPu and GTATGT consensus motifs found for Y. lipolytica (39,65) and S. cerevisiae (69), respectively. As in the Y. lipolytica genes SEC14 (39) and PYK1 (65), a 3Ј-splice acceptor CAG sequence is found one nucleotide downstream of the consensus TACTAAC box (69) or its abbreviated form CTAAC (Fig.…”

Section: Resultsmentioning

confidence: 98%

A Rac Homolog Is Required for Induction of Hyphal Growth in the Dimorphic Yeast Yarrowia lipolytica

et al. 2000

View full text Add to dashboard Cite

Dimorphism in fungi is believed to constitute a mechanism of response to adverse conditions and represents an important attribute for the development of virulence by a number of pathogenic fungal species. We have isolated YlRAC1, a gene encoding a 192-amino-acid protein that is essential for hyphal growth in the dimorphic yeast Yarrowia lipolytica and which represents the first Rac homolog described for fungi. YlRAC1 is not an essential gene, and its deletion does not affect the ability to mate or impair actin polarization in Y. lipolytica. However, strains lacking functional YlRAC1 show alterations in cell morphology, suggesting that the function of YlRAC1 may be related to some aspect of the polarization of cell growth. Northern blot analysis showed that transcription of YlRAC1 increases steadily during the yeast-to-hypha transition, while Southern blot analysis of genomic DNA suggested the presence of several RAC family members in Y. lipolytica. Interestingly, strains lacking functional YlRAC1 are still able to grow as the pseudohyphal form and to invade agar, thus pointing to a function for YlRAC1 downstream of MHY1, a previously isolated gene encoding a C 2 H 2 -type zinc finger protein with the ability to bind putative stress response elements and whose activity is essential for both hyphal and pseudohyphal growth in Y. lipolytica.

show abstract

“…The complete YlRAS2 sequence encodes a predicted protein of 249 amino acids displaying 43% identity to Ras2p and 40% identity to Ras1p of S. cerevisiae. The YlRAS2 gene contains a 348-bp intron inserted at nt 53 of the ORF, with typical Y. lipolytica splicing sites: 5Ј-GAGTAG-3Ј, 5Ј-TACTAAC-3Ј, and 5Ј-CA G-3Ј for the donor, internal, and acceptor sites, respectively (54,64). S. cerevisiae possesses two Ras homologs, RAS1 and RAS2.…”

Section: Resultsmentioning

confidence: 99%

Tagging Morphogenetic Genes by Insertional Mutagenesis in the Yeast Yarrowia lipolytica

et al. 2001

View full text Add to dashboard Cite

The yeast Yarrowia lipolytica is distantly related to Saccharomyces cerevisiae, can be genetically modified, and can grow in both haploid and diploid states in either yeast, pseudomycelial, or mycelial forms, depending on environmental conditions. Previous results have indicated that the STE and RIM pathways, which mediate cellular switching in other dimorphic yeasts, are not required for Y. lipolytica morphogenesis. To identify the pathways involved in morphogenesis, we mutagenized a wild-type strain of Y. lipolytica with a Tn3 derivative. We isolated eight tagged mutants, entirely defective in hyphal formation, from a total of 40,000 mutants and identified seven genes homologous to S. cerevisiae CDC25, RAS2, BUD6, KEX2, GPI7, SNF5, and PPH21. We analyzed their abilities to invade agar and to form pseudomycelium or hyphae under inducing conditions and their sensitivity to temperature and to Calcofluor white. Chitin staining was used to detect defects in their cell walls. Our results indicate that a functional Ras-cyclic AMP pathway is required for the formation of hyphae in Y. lipolytica and that perturbations in the processing of extracellular, possibly parietal, proteins result in morphogenetic defects.

show abstract

The isolation and characterization of the pyruvate kinase-encoding gene from the yeast Yarrowia lipolytica

Cited by 19 publications

References 17 publications

Isolation and Characterization of YlBEM1 , a Gene Required for Cell Polarization and Differentiation in the Dimorphic Yeast Yarrowia lipolytica

Isolation and Characterization of YlBEM1 , a Gene Required for Cell Polarization and Differentiation in the Dimorphic Yeast Yarrowia lipolytica

A Rac Homolog Is Required for Induction of Hyphal Growth in the Dimorphic Yeast Yarrowia lipolytica

Tagging Morphogenetic Genes by Insertional Mutagenesis in the Yeast Yarrowia lipolytica

Contact Info

Product

Resources

About