Induction of the Allantoin Degradative Enzymes in
            <i>Saccharomyces cerevisiae</i>
            by the Last Intermediate of the Pathway

Cooper, Terrance G.; Lawther, Robert P.

doi:10.1073/pnas.70.8.2340

Cited by 63 publications

(48 citation statements)

References 9 publications

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“…For DNA fiber autoradiography, cells were grown in medium containing 5 pg/ml uridine (23), pulse labeled with low specific activity [5, Ci/mM, 100 ttCi/ml ; Amersham Corp ., Arlington Heights, Ill.), washed quickly by filtration, and incubated in the same medium with fivefold higher specific activity [5,6-'Hjuracil (44.5 Ci/mM, 100 pCi/ml).…”

Section: Strains Media and Radiolabelingmentioning

confidence: 99%

Replication fork rate and origin activation during the S phase of Saccharomyces cerevisiae.

Rivin¹,

Fangman²

1980

The Journal of Cell Biology

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When the growth rate of the yeast Saccharomyces cerevisiae is limited with various nitrogen sources, the duration of the S phase is proportional to cell cycle length over a fourfold range of growth rates (C . J . Rivin and W . L . Fangman, 1980, J. Cell Biol. 85:96-107) . Molecular parameters of the S phases of these cells were examined by DNA fiber autoradiography . Changes in replication fork rate account completely for the changes in S-phase duration . No changes in origin-to-origin distances were detected . In addition, it was found that while most adjacent replication origins are activated within a few minutes of each other, new activations occur throughout the S phase .Chromosome replication occurs during an interval which can change in length at different stages of development . This variation is especially dramatic from early to late embryogenesis in some organisms (19) . The duration of the S phase can be thought of as the product of three potentially ratelimiting parameters : the rate of DNA chain elongation (fork rate), the distance between replication origins, and the time in S when different origins become active .There have been two detailed studies of the basis of S-phase length variation . In the newt Triturus the S phase of spermatocytes is one hundred times as long as that of neurula cells, and four times as long as the S phase of somatic cells (3, 4) . Fork rate does not vary in the different cell types, and the changes in S-phase length could be accounted for by changes in inter-origin spacing . Even greater S-phase variation has been observed in Drosophila in which cultured cells have a 600-min S phase while the cleavage nuclei of early embryos complete replication in <3 .5 min (1) . Again, the replication fork rate does not vary, and the observed inter-origin distance was only fourfold shorter in the early embryos . The change in S-phase duration, therefore, may reflect a change in the temporal pattern of activation of origins (or clusters of origins) . The observations that the inactive mammalian X chromosome replicates later than the active one and that the times of replication of some mammalian chromosomes exhibit tissue specificity (6, 25) also suggest that changes in the temporal pattern of origin activation can occur. In these cases, however, no change in the length of S phase has been noted .We have begun a study of the relationships of the molecular parameters of the S phase in the budding yeast Saccharomyces cerevisiae . As in other eukaryotes, DNA synthesis in yeast proceeds bidirectionally from multiple replication origins (20,21) . Estimates of the replication fork rate (21, 15) are similar to those for mammalian cells . In addition, there is evidence that the replication of specific genes and, therefore, the activation of origins occurs at specific times in the S phase (2, 17) . However, it is not known whether replication origins are activated throughout the S phase of yeast as they are in the cells of higher eukaryotes J . CELL BIOLOGY

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Section: Strains Media and Radiolabelingmentioning

confidence: 99%

Replication fork rate and origin activation during the S phase of Saccharomyces cerevisiae.

Rivin¹,

Fangman²

1980

The Journal of Cell Biology

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“…Genes encoding the allantoin-degradative system in Saccharomyces cerevisiae, for example, respond to both induction and nitrogen catabolite repression (15,25,38). The five enzyme activities of this system are present at relatively low basal levels unless compounds that can be degraded to allophanate are added to the culture medium (15). In the presence of this native inducer or its gratuitous analog oxalurate the levels of these enzymes dramatically increase (33).…”

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confidence: 99%

“…Nitrogen catabolic systems are particularly useful for such investigations, because most are subject to multiple layers of regulation. Genes encoding the allantoin-degradative system in Saccharomyces cerevisiae, for example, respond to both induction and nitrogen catabolite repression (15,25,38). The five enzyme activities of this system are present at relatively low basal levels unless compounds that can be degraded to allophanate are added to the culture medium (15).…”

mentioning

confidence: 99%

“…Nitrogen catabolic systems are particularly useful for such investigations, because most are subject to multiple layers of regulation. Genes encoding the allantoin-degradative system in Saccharomyces cerevisiae, for example, respond to both induction and nitrogen catabolite repression (15,25,38 its 3-min half-life after the addition of asparagine to the medium (12). In contrast, enzyme activity is not lost after the addition of a repressive nitrogen source, but continued enzyme synthesis ceases (12).…”

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Induction and repression of the urea amidolyase gene in Saccharomyces cerevisiae.

Genbauffe¹,

Cooper²

1986

Mol. Cell. Biol.

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The DUR1,2 gene from Saccharomyces cerevisiae has been isolated on recombinant plasmids along with all DNA between the DURI,2 and MET8 loci. DUR1,2 was found to encode a 5.7-kilobase transcript, which is consistent with our earlier suggestion that the DURi and DUR2 loci are two domains of a single multifunctional gene. Steady-state levels of the DUR1,2 transcript responded to induction and nitrogen catabolite repression in the same way as urea amidolyase activity. da)81 mutants (grown with inducer) contained barely detectable amounts of DURI,2 RNA, whereas dal80 mutants (grown without inducer) contained the same amount as a wild-type induced culture. These observations support our earlier hypothesis that DUR1,2 is transcriptionally regulated, with control being mediated by the DAL80 and DAL8I gene products. We cloned the DUR1,2_Oh mutation and found it to be a Ty insertion near sequences required for complementation of durl,2 mutations.The ROAM phenotype of the DUR1,2_Oh mutation is sharply different from that of cis-dominant, DUR80 mutations, which enhance DUR1,2 expression but do not affect the normal control pattern of the gene. There is evidence that DUR80 mutations may also be Ty insertions, which generate phenotypes that are different from those in DUR1,2-Oh mutations.An appreciation for the molecular mechanisms involved in control and integration of procaryotic metabolic pathways has been gained by studying regulons with widely differing physiological functions (28, 31). Similar information is now beginning to accumulate for eucaryotic systems (42). Nitrogen catabolic systems are particularly useful for such investigations, because most are subject to multiple layers of regulation. Genes encoding the allantoin-degradative system in Saccharomyces cerevisiae, for example, respond to both induction and nitrogen catabolite repression (15,25,38 its 3-min half-life after the addition of asparagine to the medium (12). In contrast, enzyme activity is not lost after the addition of a repressive nitrogen source, but continued enzyme synthesis ceases (12). The lack of enzyme induction does not result from inducer exclusion, because daI80 mutants, which do not require the presence of an inducer for enzyme production, are similarly devoid of allantoindegrading enzymes when grown in glucose-asparagine medium (8).Earlier kinetic studies are consistent with the hypothesis that both induction and nitrogen catabolite repression are exerted at gene expression (1-4, 16, 24-26

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“…The four degradative enzymes and at least one of the associated transport systems are inducible. Allophanate, the last pathway intermediate, and oxalurate (OXLU) function as native and gratuitous inducers, respectively (5,19). Hybridization experiments have demonstrated that, upon induction, increases in several pathway' enzymes correlate with similar increases in steadystate levels of mRNA (see Fig.…”

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cis-Dominant mutations which dramatically enhance DUR1,2 gene expression without affecting its normal regulation.

Chisholm¹,

Cooper²

1984

Mol. Cell. Biol.

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We have isolated three cis-dominant mutations which dramatically enhance DURI,2 gene expression in Saccharomyces cerevisiae. The mutant phenotype, which is expressed both in haploid and MATa/MATa diploid strains, does not appear to be an alteration of the normal control system for this gene because its expression remained fully inducible and sensitive to nitrogen catabolite repression. Instead, we found much higher levels of DURI,2-specific RNA under both uninduced and induced conditions, i.e., the overproduction trait was superimposed on normal regulation of the gene. The mutations seemed to affect gene expression in a unidirectional manner or to be specific for DURI,2 gene expression, because other genes in proximity to the mutations were not affected. We feel that these mutations may alter the chromatin structure in the vicinity of the DURJ,2 upstream control sequences or, alternatively, may be Ty insertions which no longer possess the ROAM characteristics reported by others and ourselves.

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Induction of the Allantoin Degradative Enzymes in Saccharomyces cerevisiae by the Last Intermediate of the Pathway

Cited by 63 publications

References 9 publications

Replication fork rate and origin activation during the S phase of Saccharomyces cerevisiae.

Replication fork rate and origin activation during the S phase of Saccharomyces cerevisiae.

Induction and repression of the urea amidolyase gene in Saccharomyces cerevisiae.

cis-Dominant mutations which dramatically enhance DUR1,2 gene expression without affecting its normal regulation.

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