Molecular insights on DNA delivery into Saccharomyces cerevisiae.
Understanding of the molecular system for DNA delivery into eucaryotic cells, a key to human DNA therapy, remains obscure. To understand this system, we undertook a study using the Saccharomyces cerevisiae model into which DNA delivery is easily assessed through competence (transformability) and for which all nonessential gene mutants (about 5000 strains) are available. We analyzed the competence of each of these mutants and identified three low-competence mutants, i.e., sin3Delta, she4Delta, and arc18Delta, and three high-competence mutants, i.e., pde2Delta, spf1Delta, and pmr1Delta. Through further studies using the six mutants, we concluded that the Arp2/3 activation machinery involving the Myo3/5p, Vrp1p, Las17p, Pan1p, and Arp2/3 complex is crucial to delivery (competence), and that high cAMP enhances competence via protein kinase A installing Tpk3p. We also propose that DNA is taken up via an endocytosis-like event, being at least partially different from well-known endocytosis.
Kawai
Shigeyuki
S
Department of Basic and Applied Molecular Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan.
Pham
Tuan Anh
TA
Nguyen
Ha Thu
HT
Nankai
Hirokazu
H
Utsumi
Takaaki
T
Fukuda
Yasuki
Y
Murata
Kousaku
K
D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000008:administration & dosage*; D004271:DNA, Fungal; D004705:Endocytosis; D017353:Gene Deletion; D018014:Gene Transfer Techniques; D005800:Genes, Fungal / Q000235:genetics*; D005816:Genetic Complementation Test; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D003598:Cytoskeletal Proteins; D004271:DNA, Fungal; D029701:Saccharomyces cerevisiae Proteins; D004247:DNA; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases