Cloning vectors for the synthesis of epitope-tagged, truncated and chimeric proteins in Saccharomyces cerevisiae

Foreman, Pamela K.; Davis, Ronald W.

doi:10.1016/0378-1119(94)90204-6

Cited by 56 publications

(35 citation statements)

References 17 publications

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“…The CYP76B6 ORF was excised as a XbaI/SacI or BglII fragment from pGEMT-G10H and cloned in the XbaI/SacI sites of the yeast expression vector YCpIF1 [17] or the BamHI site of plant expression vector pMOG463, respectively. The CYP76B6 derivatives and the corresponding empty vectors were introduced into Saccharomyces cerevisiae strain YPH500 [18] via the standard lithium acetate procedure, or into C. roseus cell line MP183L via particle bombardment [19].…”

Section: Expression In Transgenic Yeast and C Roseus Cellsmentioning

confidence: 99%

Geraniol 10‐hydroxylase¹, a cytochrome P450 enzyme involved in terpenoid indole alkaloid biosynthesis

Collu¹,

Unver²,

et al. 2001

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Geraniol 10-hydroxylase (G10H) is a cytochrome P450 monooxygenase involved in the biosynthesis of iridoid monoterpenoids and several classes of monoterpenoid alkaloids found in a diverse range of plant species. Catharanthus roseus (Madagascar periwinkle) contains monoterpenoid indole alkaloids, several of which are pharmaceutically important. Vinblastine and vincristine, for example, find widespread use as anticancer drugs. G10H is thought to play a key regulatory role in terpenoid indole alkaloid biosynthesis. We purified G10H from C. roseus cells. Using degenerate PCR primers based on amino acid sequence information we cloned the corresponding cDNA. The encoded CYP76B6 protein has G10H activity when expressed in C. roseus and yeast cells. The stress hormone methyljasmonate strongly induced G10h gene expression coordinately with other terpenoid indole alkaloid biosynthesis genes in a C. roseus cell culture. ß

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Section: Expression In Transgenic Yeast and C Roseus Cellsmentioning

confidence: 99%

Geraniol 10‐hydroxylase¹, a cytochrome P450 enzyme involved in terpenoid indole alkaloid biosynthesis

Collu¹,

Unver²,

et al. 2001

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“…Mutations in Ser5193Ala and Lys2013Met were made by PCR and verified by either DNA sequencing or digestion with specific restriction enzymes. The yeast expression vector YCpIF16 (P GAL1 -HA TRP1 CEN) allowed the expression of HA fusion proteins using the GAL1 promoter (12). HOG1 was cloned into the BamHI site of YCpIF16 for expression of HA-tagged HOG1.…”

Section: Yeastmentioning

confidence: 99%

Rck2 Kinase Is a Substrate for the Osmotic Stress-Activated Mitogen-Activated Protein Kinase Hog1

Bilsland-Marchesan

Ariño

Saito

et al. 2000

Molecular and Cellular Biology

136

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“…The yeast expression vector YCpIF16 (P GAL1 -hemagglutinin [HA] TRP1 ϩ CEN) allows the expression of HA fusion proteins under control of the GAL1 promoter (9), and p426TEG1 (P TEF1 -glutathione S-transferase [GST], URA3 ϩ 2m) allows the expression of GST fusion proteins under control of the P TEF1 promoter (24). Full-length and several mutant alleles of STE50 and STE11 were cloned into plasmids YCpIF16 and p426TEG1.…”

Section: Yeastmentioning

confidence: 99%

Requirement of STE50 for Osmostress-Induced Activation of the STE11 Mitogen-Activated Protein Kinase Kinase Kinase in the High-Osmolarity Glycerol Response Pathway

Posas

Witten

Saito

1998

Molecular and Cellular Biology

128

136

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Exposure of yeast cells to increases in extracellular osmolarity activates the HOG1 mitogen-activated protein (MAP) kinase cascade, which is composed of three tiers of protein kinases: (i) the SSK2, SSK22, and STE11 MAP kinase kinase kinases (MAPKKKs), (ii) the PBS2 MAPKK, and (iii) the HOG1 MAP kinase. Activation of the MAP kinase cascade is mediated by two upstream mechanisms. The SLN1-YPD1-SSK1 two-component osmosensor activates the SSK2 and SSK22 MAPKKKs by direct interaction of the SSK1 response regulator with these MAPKKKs. The second mechanism of HOG1 MAP kinase activation is independent of the twocomponent osmosensor and involves the SHO1 transmembrane protein and the STE11 MAPKKK. Only PBS2 and HOG1 are common to the two mechanisms. We conducted an exhaustive mutant screening to identify additional elements required for activation of STE11 by osmotic stress. We found that strains with mutations in the STE50 gene, in combination with ssk2⌬ ssk22⌬ mutations, were unable to induce HOG1 phosphorylation after osmotic stress. Both two-hybrid analyses and coprecipitation assays demonstrated that the N-terminal domain of STE50 binds strongly to the N-terminal domain of STE11. The binding of STE50 to STE11 is constitutive and is not affected by osmotic stress. Furthermore, the two proteins relocalize similarly after osmotic shock. It was concluded that STE50 fulfills an essential role in the activation of the high-osmolarity glycerol response pathway by acting as an integral subunit of the STE11 MAPKKK.Mitogen-activated protein (MAP) kinase cascades are common signaling modules found in both higher and lower eukaryotic cells. A typical MAP kinase cascade is composed of three tiers of protein kinases, a MAP kinase (MAPK), a MAPK kinase (MAPKK), and a MAPKK kinase (MAPKKK) (27). Yeast cells have several distinct MAP kinase cascades that transduce distinct extracellular stimuli (e.g., mating pheromone, high osmolarity, low osmolarity, and nitrogen starvation) (10, 19). Budding yeast (Saccharomyces cerevisiae) responds to increases in osmolarity in the extracellular environment by activating the HOG1 MAP kinase cascade. This cascade is essential for the survival of yeast in high-osmolarity environments (4, 5). Because a major outcome of the activation of this MAPK pathway is the elevated synthesis of glycerol, this pathway is referred to as the HOG (high-osmolarity glycerol response) pathway (2, 5). Extracellular hyperosmolarity is detected by either of two transmembrane proteins, SLN1 and SHO1 (Fig. 1).SLN1 is a part of a complex regulatory system with homology to prokaryotic two-component signal transducers. The yeast two-component osmosensor is composed of SLN1, a transmembrane protein that contains an extracellular sensor domain and cytoplasmic histidine kinase and receiver domains, the intermediary protein YPD1, and the response regulator SSK1 (14,15,20). The SLN1-YPD1-SSK1 two-component osmosensor works by a multistep phosphorelay mechanism (20). The unphosphorylated form of SSK1 activates SSK2 and SSK22 MAPKK...

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Cloning vectors for the synthesis of epitope-tagged, truncated and chimeric proteins in Saccharomyces cerevisiae

Cited by 56 publications

References 17 publications

Geraniol 10‐hydroxylase¹, a cytochrome P450 enzyme involved in terpenoid indole alkaloid biosynthesis

Geraniol 10‐hydroxylase¹, a cytochrome P450 enzyme involved in terpenoid indole alkaloid biosynthesis

Rck2 Kinase Is a Substrate for the Osmotic Stress-Activated Mitogen-Activated Protein Kinase Hog1

Requirement of STE50 for Osmostress-Induced Activation of the STE11 Mitogen-Activated Protein Kinase Kinase Kinase in the High-Osmolarity Glycerol Response Pathway

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Cloning vectors for the synthesis of epitope-tagged, truncated and chimeric proteins in Saccharomyces cerevisiae

Cited by 56 publications

References 17 publications

Geraniol 10‐hydroxylase1, a cytochrome P450 enzyme involved in terpenoid indole alkaloid biosynthesis

Geraniol 10‐hydroxylase1, a cytochrome P450 enzyme involved in terpenoid indole alkaloid biosynthesis

Rck2 Kinase Is a Substrate for the Osmotic Stress-Activated Mitogen-Activated Protein Kinase Hog1

Requirement of STE50 for Osmostress-Induced Activation of the STE11 Mitogen-Activated Protein Kinase Kinase Kinase in the High-Osmolarity Glycerol Response Pathway

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Geraniol 10‐hydroxylase¹, a cytochrome P450 enzyme involved in terpenoid indole alkaloid biosynthesis

Geraniol 10‐hydroxylase¹, a cytochrome P450 enzyme involved in terpenoid indole alkaloid biosynthesis