Limits...
Genome-wide screening for genes whose deletions confer sensitivity to mutagenic purine base analogs in yeast.

Stepchenkova EI, Kozmin SG, Alenin VV, Pavlov YI - BMC Genet. (2005)

Bottom Line: We screened the library of yeast deletion mutants for sensitivity to the toxic and mutagenic action of HAP and AHA.We developed a method for screening the yeast deletion library for sensitivity to the mutagenic and toxic action of base analogs and identified 16 novel genes controlling pathways of protection from HAP.Three of them also protect from AHA.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Genetics, Sankt-Petersburg State University, Sankt-Petersburg, 199034, Russia. stepchenkova@yahoo.com <stepchenkova@yahoo.com>

ABSTRACT

Background: N-hydroxylated base analogs, such as 6-hydroxylaminopurine (HAP) and 2-amino-6-hydroxylaminopurine (AHA), are strong mutagens in various organisms due to their ambiguous base-pairing properties. The systems protecting cells from HAP and related noncanonical purines in Escherichia coli include specialized deoxyribonucleoside triphosphatase RdgB, DNA repair endonuclease V, and a molybdenum cofactor-dependent system. Fewer HAP-detoxification systems have been identified in yeast Saccharomyces cerevisiae and other eukaryotes. Cellular systems protecting from AHA are unknown. In the present study, we performed a genome-wide search for genes whose deletions confer sensitivity to HAP and AHA in yeast.

Results: We screened the library of yeast deletion mutants for sensitivity to the toxic and mutagenic action of HAP and AHA. We identified novel genes involved in the genetic control of base analogs sensitivity, including genes controlling purine metabolism, cytoskeleton organization, and amino acid metabolism.

Conclusion: We developed a method for screening the yeast deletion library for sensitivity to the mutagenic and toxic action of base analogs and identified 16 novel genes controlling pathways of protection from HAP. Three of them also protect from AHA.

Show MeSH

Related in: MedlinePlus

Scheme of the protocol for screening the yeast deletions library for base analog sensitivity and induced mutagenesis.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC1173102&req=5

Figure 2: Scheme of the protocol for screening the yeast deletions library for base analog sensitivity and induced mutagenesis.

Mentions: To develop a useful method for searching the yeast mutants sensitive to base analogs, we calibrated the experimental conditions using the wild-type strain, BY4742, and two previously described HAP-sensitive mutants, ham1 and aah1 (see [6,16]), created on BY4742 background. As shown in Fig. 2 and described in Materials and Methods, yeast were grown in a 96-well microtiter plate and then transferred, using a multiprong replicator device, to YPD plates containing base analogs. An induction of the Canr mutants was monitored by replica-plating to the minimal-medium plates containing L-canavanine.


Genome-wide screening for genes whose deletions confer sensitivity to mutagenic purine base analogs in yeast.

Stepchenkova EI, Kozmin SG, Alenin VV, Pavlov YI - BMC Genet. (2005)

Scheme of the protocol for screening the yeast deletions library for base analog sensitivity and induced mutagenesis.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC1173102&req=5

Figure 2: Scheme of the protocol for screening the yeast deletions library for base analog sensitivity and induced mutagenesis.
Mentions: To develop a useful method for searching the yeast mutants sensitive to base analogs, we calibrated the experimental conditions using the wild-type strain, BY4742, and two previously described HAP-sensitive mutants, ham1 and aah1 (see [6,16]), created on BY4742 background. As shown in Fig. 2 and described in Materials and Methods, yeast were grown in a 96-well microtiter plate and then transferred, using a multiprong replicator device, to YPD plates containing base analogs. An induction of the Canr mutants was monitored by replica-plating to the minimal-medium plates containing L-canavanine.

Bottom Line: We screened the library of yeast deletion mutants for sensitivity to the toxic and mutagenic action of HAP and AHA.We developed a method for screening the yeast deletion library for sensitivity to the mutagenic and toxic action of base analogs and identified 16 novel genes controlling pathways of protection from HAP.Three of them also protect from AHA.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Genetics, Sankt-Petersburg State University, Sankt-Petersburg, 199034, Russia. stepchenkova@yahoo.com <stepchenkova@yahoo.com>

ABSTRACT

Background: N-hydroxylated base analogs, such as 6-hydroxylaminopurine (HAP) and 2-amino-6-hydroxylaminopurine (AHA), are strong mutagens in various organisms due to their ambiguous base-pairing properties. The systems protecting cells from HAP and related noncanonical purines in Escherichia coli include specialized deoxyribonucleoside triphosphatase RdgB, DNA repair endonuclease V, and a molybdenum cofactor-dependent system. Fewer HAP-detoxification systems have been identified in yeast Saccharomyces cerevisiae and other eukaryotes. Cellular systems protecting from AHA are unknown. In the present study, we performed a genome-wide search for genes whose deletions confer sensitivity to HAP and AHA in yeast.

Results: We screened the library of yeast deletion mutants for sensitivity to the toxic and mutagenic action of HAP and AHA. We identified novel genes involved in the genetic control of base analogs sensitivity, including genes controlling purine metabolism, cytoskeleton organization, and amino acid metabolism.

Conclusion: We developed a method for screening the yeast deletion library for sensitivity to the mutagenic and toxic action of base analogs and identified 16 novel genes controlling pathways of protection from HAP. Three of them also protect from AHA.

Show MeSH
Related in: MedlinePlus