Expression of Saccharomyces adenylate kinase gene in Candida boidinii under the regulation of its alcohol oxidase promoter

1995 ◽  
Vol 42 (6) ◽  
pp. 860-864 ◽  
Author(s):  
Y. Sakai ◽  
T. Rogi ◽  
R. Takeuchi ◽  
N. Kato ◽  
Y. Tani
Keyword(s):  
Adenylate Kinase ◽  
Alcohol Oxidase ◽  
Candida Boidinii ◽  
Kinase Gene ◽  
Alcohol Oxidase Promoter

scholarly journals Peroxisomal assembly: membrane proliferation precedes the induction of the abundant matrix proteins in the methylotrophic yeast Candida boidinii

1990 ◽  
Vol 96 (4) ◽  
pp. 583-590
Author(s):  
M. Veenhuis ◽  
J.M. Goodman
Keyword(s):  
Early Stage ◽  
Morphological Changes ◽  
Polyclonal Antibodies ◽  
Alcohol Oxidase ◽  
Methylotrophic Yeast ◽  
Candida Boidinii ◽  
Peroxisomal Membrane ◽  
Cell Induction ◽  
The Matrix ◽  
Dihydroxyacetone Synthase

Peroxisomes are massively induced when methylotrophic yeasts are cultured in medium containing methanol. These organelles contain enzymes that catalyze the initial steps of methanol assimilation. In Candida boidinii, a methylotrophic yeast, the peroxisomal matrix (internal compartment) is composed almost exclusively of two proteins, alcohol oxidase and dihydroxyacetone synthase; catalase is present in much lower abundance. Monoclonal and polyclonal antibodies are available against peroxisomal matrix and membrane proteins. These were utilized to correlate the induction of specific proteins with the morphological changes occurring during peroxisomal proliferation. Cells cultured in glucose-containing medium contain two to five small microbodies, which are identifiable by catalase staining and immunoreactivity with a monoclonal antibody against PMP47, an integral peroxisomal membrane protein. Three stages of proliferation can be distinguished when cells are switched to methanol as the carbon source. (1) There is an early stage (within 1 h) in which several peroxisomes develop from a preexisting organelle. This is accompanied by an increase in catalase activity and an induction of PMP47, but no detectable induction of alcohol oxidase or dihydroxyacetone synthase is observed. (2) From 1 to 2.5 h there is further division of these microbodies until up to 30 small peroxisomes generally are present in each of one or two clusters per cell. Induction of alcohol oxidase, dihydroxyacetone synthase and PMP20, a protein that is distributed in the matrix and membrane, is detectable during this time. Serial sections reveal that some peroxisomes remain uninduced while others undergo proliferation. Such sections also show no obvious connections between peroxisomes within clusters.(ABSTRACT TRUNCATED AT 250 WORDS)

An expanded adenylate kinase gene family in the protozoan parasite Trypanosoma cruzi

2006 ◽  
Vol 1760 (6) ◽  
pp. 913-921 ◽  
Author(s):  
Leon A. Bouvier ◽  
Mariana R. Miranda ◽  
Gaspar E. Canepa ◽  
Maria Júlia M. Alves ◽  
Claudio A. Pereira
Keyword(s):  
Trypanosoma Cruzi ◽  
Gene Family ◽  
Adenylate Kinase ◽  
Protozoan Parasite ◽  
Kinase Gene

scholarly journals Intracellular Positioning of Isoforms Explains an Unusually Large Adenylate Kinase Gene Family in the Parasite Trypanosoma brucei

2005 ◽  
Vol 280 (12) ◽  
pp. 11781-11789 ◽  
Author(s):  
Michael L. Ginger ◽  
E. Solange Ngazoa ◽  
Claudio A. Pereira ◽  
Timothy J. Pullen ◽  
Mostafa Kabiri ◽  
...  
Keyword(s):  
Gene Family ◽  
Trypanosoma Brucei ◽  
Adenylate Kinase ◽  
Kinase Gene

scholarly journals The absence of Pmp47, a putative yeast peroxisomal transporter, causes a defect in transport and folding of a specific matrix enzyme.

1996 ◽  
Vol 134 (1) ◽  
pp. 37-51 ◽  
Author(s):  
Y Sakai ◽  
A Saiganji ◽  
H Yurimoto ◽  
K Takabe ◽  
H Saiki ◽  
...  
Keyword(s):  
Single Gene ◽  
Alcohol Oxidase ◽  
Candida Boidinii ◽  
High Electron ◽  
Gene Encoding ◽  
Peroxisomal Membrane ◽  
Carboxyl Terminal ◽  
Dihydroxyacetone Synthase ◽  
Physiological And Biochemical ◽  
Solute Transporters

Candida boidinii Pmp47, an integral peroxisomal membrane protein, belongs to a family of mitochondrial solute transporters (e.g., ATP/ADP exchanger), and is the only known peroxisomal member of this family. However, its physiological and biochemical functions have been unrevealed because of the difficulties in the molecular genetics of C. boidinii. In this study, we first isolated the PMP47 gene, which was the single gene encoding for Pmp47 in a gene-engineerable strain S2 of C. boidinii. Sequence analysis revealed that it was very similar to PMP47A and PMP47B genes from a polyploidal C. Boidinii strain (ATCC32195). Next, the PMP47 gene was disrupted and the disruption strain (pmp47delta) was analyzed. Depletion of PMP47 from strain S2 resulted in a retarded growth on oleate and a complete loss of growth on methanol. Both growth substrates require peroxisomal metabolism. EM observations revealed the presence of peroxisomes in methanol- and oleate-induced cells of pmp47delta, but in reduced numbers, and the presence of material of high electron density in the cytoplasm in both cases. Methanol-induced cells of pmp47delta were investigated in detail. The activity of one of the methanol-induced peroxisome matrix enzymes, dihydroxyacetone synthase (DHAS), was not detected in pmp47delta. Further biochemical and immunocytochemical experiments revealed that the DHAS protein aggregated in the cytoplasm as an inclusion body, while two other peroxisome matrix enzymes, alcohol oxidase (AOD) and catalase, were active and found in peroxisomes. Two peroxisome-deficient mutants, strains M6 and M13 (described in previous studies), retained DHAS activity although it was mislocalized to the cytoplasm and the nucleus. We disrupted PMP47 in these peroxisome-deficient mutants. In both strains, M6-pmp47delta and M13-pmp47delta, DHAS was enzymatically active and was located in the cytoplasm and the nucleus. We suggest that an unknown small molecule, which PMP47 transports, is necessary for the folding or the translocation machinery of DHAS within peroxisomes. Pmp47 does not catalyze folding directly because active DHAS is observed in the M6-pmp47delta and M13-pmp47delta strains. Since both AOD and DHAS have the PTS1 motif sequences at their carboxyl terminal, our results first show that depletion of Pmp47 could dissect the peroxisomal import pathway (PTS1 pathway) of these proteins.

scholarly journals Relationship between bacterial virulence and nucleotide metabolism: a mutation in the adenylate kinase gene renders Yersinia pestis avirulent

2003 ◽  
Vol 373 (2) ◽  
pp. 515-522 ◽  
Author(s):  
Hélène MUNIER-LEHMANN ◽  
Viviane CHENAL-FRANCISQUE ◽  
Mihaela IONESCU ◽  
Petya CHRISTOVA ◽  
Jeannine FOULON ◽  
...  
Keyword(s):  
Yersinia Pestis ◽  
Adenylate Kinase ◽  
Biochemical Properties ◽  
Nucleotide Metabolism ◽  
Growth Defect ◽  
Permissive Temperature ◽  
Wild Type ◽  
Kinase Gene ◽  
E Coli ◽  
Colony Forming Units

Nucleoside monophosphate kinases (NMPKs) are essential catalysts for bacterial growth and multiplication. These enzymes display high primary sequence identities among members of the family Enterobacteriaceae. Yersinia pestis, the causative agent of plague, belongs to this family. However, it was previously shown that its thymidylate kinase (TMPKyp) exhibits biochemical properties significantly different from those of its Escherichia coli counterpart [Chenal-Francisque, Tourneux, Carniel, Christova, Li de la Sierra, Barzu and Gilles (1999) Eur. J. Biochem. 265, 112–119]. In this work, the adenylate kinase (AK) of Y. pestis (AKyp) was characterized. As with TMPKyp, AKyp displayed a lower thermodynamic stability than other studied AKs. Two mutations in AK (Ser129→Phe and Pro87→Ser), previously shown to induce a thermosensitive growth defect in E. coli, were introduced into AKyp. The recombinant variants had a lower stability than wild-type AKyp and a higher susceptibility to proteolytic digestion. When the Pro87→Ser substitution was introduced into the chromosomal adk gene of Y. pestis, growth of the mutant strain was altered at the non-permissive temperature of 37 °C. In virulence testings, less than 50 colony forming units (CFU) of wild-type Y. pestis killed 100% of the mice upon subcutaneous infection, whereas bacterial loads as high as 1.5×104 CFU of the adk mutant were unable to kill any animals.

scholarly journals Cloning and sequencing of the adenylate kinase gene (adk) ofEscherichia coli

1985 ◽  
Vol 13 (19) ◽  
pp. 7139-7151 ◽  
Author(s):  
M. Brune ◽  
R. Schumann ◽  
F. Wittinghofer
Keyword(s):  
Adenylate Kinase ◽  
Ofescherichia Coli ◽  
Cloning And Sequencing ◽  
Kinase Gene
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