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1.
Mol Genet Genomics ; 274(5): 528-38, 2005 Dec.
Article in English | MEDLINE | ID: mdl-16231152

ABSTRACT

In Saccharomyces cerevisiae, HOT1-stimulated recombination has been implicated in maintaining homology between repeated ribosomal RNA genes. The ability of HOT1 to stimulate genetic exchange requires RNA polymerase I transcription across the recombining sequences. The trans-acting nuclear mutation hrm3-1 specifically reduces HOT1-dependent recombination and prevents cell growth at 37 degrees . The HRM3 gene is identical to DEG1. Excisive, but not gene replacement, recombination is reduced in HOT1-adjacent sequences in deg1Delta mutants. Excisive recombination within the genomic rDNA repeats is also decreased. The hypo-recombination and temperature-sensitive phenotypes of deg1Delta mutants are recessive. Deletion of DEG1 did not affect the rate of transcription from HOT1 or rDNA suggesting that while transcription is necessary it is not sufficient for HOT1 activity. Pseudouridine synthase 3 (Pus3p), the DEG1 gene product, modifies the anticodon arm of transfer RNA at positions 38 and 39 by catalyzing the conversion of uridine to pseudouridine. Cells deficient in pseudouridine synthases encoded by PUS1, PUS2 or PUS4 displayed no recombination defects, indicating that Pus3p plays a specific role in HOT1 activity. Pus3p is unique in its ability to modulate frameshifting and readthrough events during translation, and this aspect of its activity may be responsible for HOT1 recombination phenotypes observed in deg1 mutants.


Subject(s)
Recombination, Genetic/physiology , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae Proteins/physiology , Saccharomyces cerevisiae/genetics , Transcription Factors/physiology , Alleles , Base Sequence , Blotting, Western , Cloning, Molecular , DNA Primers , Epitopes/genetics , Gene Deletion , Genes, Fungal , Intramolecular Transferases
2.
Mol Genet Genomics ; 272(3): 264-74, 2004 Oct.
Article in English | MEDLINE | ID: mdl-15349770

ABSTRACT

HOT1 is a mitotic recombination hotspot derived from yeast rDNA. To further study HOT1 function, trans-acting H OT1 recombination mutants (hrm) that alter hotspot activity were isolated. hrm2-1 mutants have decreased HOT1 activity and grow slowly. The HRM2 gene was cloned and found to be identical to SCH9, a gene that affects a growth-control mechanism that is partially redundant with the cAMP-dependent protein kinase A (PKA) pathway. Deletion of SCH9 decreases HOT1 and rDNA recombination but not other mitotic exchange. Although high levels of RNA polymerase I transcription initiated at HOT1 are required for its recombination-stimulating activity, sch9 mutations do not affect transcription initiated within HOT1. Thus, transcription is necessary but not sufficient for HOT1 activity. TPK1, which encodes a catalytic subunit of PKA, is a multicopy suppressor of the recombination and growth defects of sch9 mutants, suggesting that increased PKA activity compensates for SCH9 loss. RAS2( val19), which codes for a hyperactive RAS protein and increases PKA activity, suppresses both phenotypic defects of sch9 mutants. In contrast to TPK1 and RAS2(val19), the gene for split zinc finger protein 1 (SFP1) on a multicopy vector suppresses only the growth defects of sch9 mutants, indicating that growth and HOT1 functions of Sch9p are separable. Sch9p may affect signal transduction pathways which regulate proteins that are specifically required for HOT1-stimulated exchange.


Subject(s)
Protein Kinases/physiology , Recombination, Genetic/physiology , Saccharomyces cerevisiae Proteins/physiology , Saccharomyces cerevisiae/enzymology , Transcription Factors/physiology , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/growth & development
3.
Mol Cell Biol ; 20(13): 4948-57, 2000 Jul.
Article in English | MEDLINE | ID: mdl-10848619

ABSTRACT

In the ribosomal DNA of Saccharomyces cerevisiae, sequences in the nontranscribed spacer 3' of the 35S ribosomal RNA gene are important to the polar arrest of replication forks at a site called the replication fork barrier (RFB) and also to the cis-acting, mitotic hyperrecombination site called HOT1. We have found that the RFB and HOT1 activity share some but not all of their essential sequences. Many of the mutations that reduce HOT1 recombination also decrease or eliminate fork arrest at one of two closely spaced RFB sites, RFB1 and RFB2. A simple model for the juxtaposition of RFB and HOT1 sequences is that the breakage of strands in replication forks arrested at RFB stimulates recombination. Contrary to this model, we show here that HOT1-stimulated recombination does not require the arrest of forks at the RFB. Therefore, while HOT1 activity is independent of replication fork arrest, HOT1 and RFB require some common sequences, suggesting the existence of a common trans-acting factor(s).


Subject(s)
DNA Replication , DNA, Ribosomal/chemistry , Recombination, Genetic , Saccharomyces cerevisiae Proteins , Transcription Factors/genetics , DNA, Fungal/chemistry , DNA, Fungal/genetics , DNA, Fungal/metabolism , DNA, Ribosomal/genetics , DNA, Ribosomal/metabolism , Deoxyribonuclease HindIII/genetics , Deoxyribonuclease HindIII/metabolism , Deoxyribonucleases, Type II Site-Specific/genetics , Deoxyribonucleases, Type II Site-Specific/metabolism , Mutation , Saccharomyces cerevisiae/genetics
4.
Mol Cell Biol ; 19(12): 8254-62, 1999 Dec.
Article in English | MEDLINE | ID: mdl-10567550

ABSTRACT

To investigate the mechanism of action of volatile anesthetics, we are studying mutants of the yeast Saccharomyces cerevisiae that have altered sensitivity to isoflurane, a widely used clinical anesthetic. Several lines of evidence from these studies implicate a role for ubiquitin metabolism in cellular response to volatile anesthetics: (i) mutations in the ZZZ1 gene render cells resistant to isoflurane, and the ZZZ1 gene is identical to BUL1 (binds ubiquitin ligase), which appears to be involved in the ubiquitination pathway; (ii) ZZZ4, which we previously found is involved in anesthetic response, is identical to the DOA1/UFD3 gene, which was identified based on altered degradation of ubiquitinated proteins; (iii) analysis of zzz1Delta zzz4Delta double mutants suggests that these genes encode products involved in the same pathway for anesthetic response since the double mutant is no more resistant to anesthetic than either of the single mutant parents; (iv) ubiquitin ligase (MDP1/RSP5) mutants are altered in their response to isoflurane; and (v) mutants with decreased proteasome activity are resistant to isoflurane. The ZZZ1 and MDP1/RSP5 gene products appear to play important roles in determining effective anesthetic dose in yeast since increased levels of either gene increases isoflurane sensitivity whereas decreased activity decreases sensitivity. Like zzz4 strains, zzz1 mutants are resistant to all five volatile anesthetics tested, suggesting there are similarities in the mechanisms of action of a variety of volatile anesthetics in yeast and that ubiquitin metabolism affects response to all the agents examined.


Subject(s)
Adaptor Proteins, Signal Transducing , Anesthetics, Inhalation/pharmacology , Carrier Proteins/metabolism , Isoflurane/pharmacology , Ligases/metabolism , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/metabolism , Ubiquitin-Protein Ligase Complexes , Ubiquitins/metabolism , Carrier Proteins/genetics , Cysteine Endopeptidases , Drug Resistance, Microbial , Endosomal Sorting Complexes Required for Transport , Enflurane/pharmacology , Fungal Proteins/genetics , Fungal Proteins/metabolism , Gene Expression , Halothane/pharmacology , Ligases/genetics , Methoxyflurane/pharmacology , Methyl Ethers/pharmacology , Multienzyme Complexes , Mutagenesis , Phenotype , Proteasome Endopeptidase Complex , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/growth & development , Sevoflurane , Temperature , Ubiquitin-Protein Ligases
5.
Mol Cell ; 3(4): 447-55, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10230397

ABSTRACT

A cause of aging in yeast is the accumulation of circular species of ribosomal DNA (rDNA) arising from the 100-200 tandemly repeated copies in the genome. We show here that mutation of the FOB1 gene slows the generation of these circles and thus extends life span. Fob1p is known to create a unidirectional block to replication forks in the rDNA. We show that Fob1p is a nucleolar protein, suggesting a direct involvement in the replication fork block. We propose that this block can trigger aging by causing chromosomal breaks, the repair of which results in the generation of rDNA circles. These findings may provide a novel link between metabolic rate and aging in yeast and, perhaps, higher organisms.


Subject(s)
DNA Replication/genetics , DNA-Binding Proteins , Fungal Proteins/genetics , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/genetics , DNA Repair/genetics , DNA, Circular/genetics , DNA, Ribosomal/genetics , Fluorescent Antibody Technique , Gene Deletion , Green Fluorescent Proteins , Luminescent Proteins , Nuclear Proteins/genetics , Repetitive Sequences, Nucleic Acid
6.
Anesthesiology ; 89(1): 174-81, 1998 Jul.
Article in English | MEDLINE | ID: mdl-9667307

ABSTRACT

BACKGROUND: In animals, combinations of volatile anesthetics are additive for inducing anesthesia. Furthermore, although there is a correlation between lipophilicity and anesthetic potency, not all volatile lipophilic compounds are anesthetic. Previously the authors demonstrated the effects of volatile anesthetics on the eukaryote Saccharomyces cerevisiae (yeast). To further relate anesthetic action in this organism to mammals, anesthetic additivity and effects of volatile, lipophilic nonanesthetics were studied. In addition, yeast pleiotropic drug-resistance (Pdr) mutants, which confer resistance to various lipophilic compounds, were tested to determine if they are involved in anesthetic response. METHODS: Yeast strains were grown to saturation in liquid culture, diluted, plated on various solid media, incubated, and scored for growth. RESULTS: Combinations of volatile anesthetics inhibit growth of wild-type (Zzz+) but not anesthetic-resistant (Zzz-) strains when additive concentrations equal 1 minimum inhibitory concentration (MIC). Two volatile, lipophilic compounds that are nonanesthetic in mammals do not inhibit yeast growth. Zzz- mutants remain sensitive to drugs used to identify yeast PDR genes. Conversely Pdr strains, which are resistant to various lipophilic compounds, remain sensitive to volatile anesthetics. CONCLUSIONS: Yeast growth is inhibited in an additive manner by volatile anesthetics. Volatile, lipophilic compounds devoid of anesthetic activity in mammals do not inhibit yeast growth. Zzz- mutants appear to be specifically resistant to volatile anesthetics and distinct from known Pdr mutants. These results suggest that volatile anesthetics behave in a parallel manner in yeast and mammals, making yeast a useful model to investigate the molecular effects of these compounds in living cells.


Subject(s)
Anesthetics, Inhalation/pharmacology , Isoflurane/pharmacology , Methoxyflurane/pharmacology , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/physiology , Drug Resistance, Multiple/genetics , Mutation
7.
Biochem J ; 331 ( Pt 1): 193-9, 1998 Apr 01.
Article in English | MEDLINE | ID: mdl-9512479

ABSTRACT

A number of cDNA fragments coding for bovine submaxillary mucin (BSM) were cloned, and the nucleotide sequence of the largest clone, BSM421, was determined. Two peptide sequences determined from the purified apoBSM were found near the N-terminus of the mucin-coding region of BSM421. This clone does not contain a start or stop codon, but its 3' end overlaps with the 5' end of a previously isolated clone, lambdaBSM10. The composite sequence of 1589 amino acid residues consists of five distinct protein domains, which are numbered from the C-terminus. The cysteine-rich domain I can be further divided into a von Willebrand factor type C repeat and a cystine knot. Domains III and V consist of similar repeated peptide sequences with an average of 47 residues. Domains II and IV do not contain such sequences but are similar to domains III and V in being rich in serine and threonine, many of which are predicted to be potential O-glycosylation sites. Domain III also contains two sequences that match the ATP/GTP-binding site motif A (P-loop). Only beta-strands and no alpha-helices are predicted for the partial deduced amino acid sequence. Northern analysis of submaxillary gland RNA with the BSM421 probe detected multiple messages of BSM with sizes from 1.1 to over 10 kb. The tandemly repeated, non-identical peptide sequences of approx. 47 residues in domains III and V of BSM differ from the tandemly repeated, identical 81-residue sequences of pig submaxillary mucin (PSM), although both BSM and PSM contain similar C-terminal domains. In contrast, two peptide sequences of ovine submaxillary mucin are highly similar (86% and 65% identical respectively) to the corresponding sequences in domain V of BSM.


Subject(s)
Mucins/metabolism , Peptides/genetics , Submandibular Gland/metabolism , Amino Acid Sequence , Animals , Base Sequence , Cattle , Cloning, Molecular , DNA, Complementary/analysis , Molecular Sequence Data , Repetitive Sequences, Nucleic Acid , Sequence Analysis
8.
Mol Cell Biol ; 16(7): 3446-53, 1996 Jul.
Article in English | MEDLINE | ID: mdl-8668160

ABSTRACT

The mechanism(s) and site(s) of action of volatile inhaled anesthetics are unknown in spite of the clinical use of these agents for more than 150 years. In the present study, the model eukaryote Saccharomyces cerevisiae was used to investigate the action of anesthetic agents because of its powerful molecular genetics. It was found that growth of yeast cells is inhibited by the five common volatile anesthetics tested (isoflurane, halothane, enflurane, sevoflurane, and methoxyflurane). Growth inhibition by the agents is relatively rapid and reversible. The potency of these compounds as yeast growth inhibitors directly correlates with their lipophilicity as is predicted by the Meyer-Overton relationship, which directly correlates anesthetic potency of agents and their lipophilicity. The effects of isoflurane on yeast cells were characterized in the most detail. Yeast cells survive at least 48 h in a concentration of isoflurane that inhibits colony formation. Mutants resistant to the growth-inhibitory effects of isoflurane are readily selected. The gene identified by one of these mutations, zzz4-1, has been cloned and characterized. The predicted ZZZ4 gene product has extensive homology to phospholipase A2-activating protein, a GO effector protein of mice. Both zzz4-1 and a deletion of ZZZ4 confer resistance to all five of the agents tested, suggesting that signal transduction may be involved in the response of these cells to volatile anesthetics.


Subject(s)
Anesthetics, Inhalation/pharmacology , Carrier Proteins , Genes, Fungal , Methyl Ethers , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/drug effects , Adaptor Proteins, Signal Transducing , Amino Acid Sequence , Animals , Base Sequence , DNA, Fungal/chemistry , DNA, Fungal/drug effects , DNA, Fungal/genetics , Enflurane/pharmacology , Ethers/pharmacology , Fungal Proteins/chemistry , Fungal Proteins/genetics , Halothane/pharmacology , Isoflurane/pharmacology , Methoxyflurane/pharmacology , Mice , Molecular Sequence Data , Mutagenesis , Oligodeoxyribonucleotides , Proteins/chemistry , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/growth & development , Sequence Homology, Amino Acid , Sevoflurane , Structure-Activity Relationship
9.
Genetics ; 141(3): 845-55, 1995 Nov.
Article in English | MEDLINE | ID: mdl-8582631

ABSTRACT

When inserted at novel locations in the yeast genome, the Saccharomyces cerevisiae recombination hotspot HOT1 stimulates mitotic exchange in adjacent sequences. HOT1 is derived from the rDNA repeat unit, and the sequences required for the recombination-stimulatory activity closely correspond to the rDNA transcription enhancer and initiation site, suggesting there is an association between high levels of RNA polymerase I transcription and increased recombination. To directly test whether RNA polymerase I is essential for HOT1 activity, a subunit of RNA polymerase I was deleted in a strain in which rRNA is transcribed by RNA polymerase II. HOT1 is completely inactive in this strain. Deletion analysis and site-directed mutagenesis were used to further define the sequences within the rDNA enhancer required for HOT1 activity. These studies show that the enhancer contains at least four distinct regions that are required for hotspot activity. In most cases mutations in these regions also decrease transcription from this element, further confirming the association of recombination and transcription.


Subject(s)
DNA, Fungal/genetics , DNA, Ribosomal/genetics , Fungal Proteins/metabolism , RNA Polymerase I/metabolism , Recombination, Genetic , Saccharomyces cerevisiae/genetics , Base Sequence , Enhancer Elements, Genetic , Mitosis , Molecular Sequence Data , Mutagenesis, Site-Directed , RNA Polymerase I/deficiency , RNA Polymerase II/metabolism , Saccharomyces cerevisiae/enzymology , Sequence Alignment , Sequence Deletion , Sequence Homology, Nucleic Acid , Transcription, Genetic
10.
Genetics ; 135(3): 711-8, 1993 Nov.
Article in English | MEDLINE | ID: mdl-8293975

ABSTRACT

The preservation of sequence homogeneity and copy number of tandemly repeated genes may require specific mechanisms or regulation of recombination. We have identified mutations that specifically affect recombination among natural repetitions in the yeast Saccharomyces cerevisiae. The rrm3 mutation stimulates mitotic recombination in the naturally occurring tandem repeats of the rDNA and copper chelatin (CUP1) genes. This mutation does not affect recombination of several other types of repeated genes tested including Ty elements, mating type information and duplications created by transformation. In addition to stimulating exchange among the multiple CUP1 repeats at their natural chromosomal location, rrm3 also increases recombination of a duplication of CUP1 units present at his4. This suggests that the RRM3 gene may encode a sequence-specific factor that contributes to a global suppression of mitotic exchange in sequences that can be maintained as tandem arrays.


Subject(s)
Genes, Fungal , Genes, Mating Type, Fungal , Multigene Family , Recombination, Genetic , Saccharomyces cerevisiae/genetics , Biological Evolution , DNA, Fungal/genetics , DNA, Ribosomal/genetics , Genetic Complementation Test , Metalloproteins/genetics
11.
Genetics ; 127(1): 31-8, 1991 Jan.
Article in English | MEDLINE | ID: mdl-2016045

ABSTRACT

HOT1 is a cis-acting recombination-stimulatory sequence isolated from the rDNA repeat unit of yeast. The ability of HOT1 to stimulate mitotic exchange appears to depend on its ability to promote high levels of RNA polymerase I transcription. A qualitative colony color sectoring assay was developed to screen for trans-acting mutations that alter the activity of HOT1. Both hypo-recombination and hyper-recombination mutants were isolated. Genetic analysis of seven HOT1 recombination mutants (hrm) that decrease HOT1 activity shows that they behave as recessive nuclear mutations and belong to five linkage groups. Three of these mutations, hrm1, hrm2, and hrm3, also decrease rDNA exchange but do not alter recombination in the absence of HOT1. Another mutation, hrm4, decreases HOT1-stimulated recombination but does not affect rDNA recombination or exchange in the absence of HOT1. Two new alleles of RAD52 were also isolated using this screen. With regard to HOT1 activity, rad52 is epistatic to all four hrm mutations indicating that the products of the HRM genes and of RAD52 mediate steps in the same recombination pathway. Finding mutations that decrease both the activity of HOT1 and exchange in the rDNA supports the hypothesis that HOT1 plays a role in rDNA recombination.


Subject(s)
DNA, Fungal/genetics , DNA, Ribosomal/genetics , Mutation , RNA Polymerase I/metabolism , Recombination, Genetic/genetics , Saccharomyces cerevisiae/genetics , Base Sequence , Epistasis, Genetic , Mutagenesis/radiation effects , Plasmids , Repetitive Sequences, Nucleic Acid , Ultraviolet Rays
12.
Genetics ; 126(1): 41-52, 1990 Sep.
Article in English | MEDLINE | ID: mdl-2227387

ABSTRACT

We examined the genetic control of the activity of HOT1, a cis-acting recombination-stimulatory sequence of Saccharomyces cerevisiae. Mutations in RAD1 and RAD52 decrease the ability of HOT1 to stimulate intrachromosomal recombination while mutations in RAD4 and RAD50 do not affect HOT1 activity. In rad1 delta strains, the stimulation of excisive recombination by HOT1 is decreased while the rate of gene replacement is not affected. In rad52-8 strains the ability of HOT1 to stimulate both excisive recombination and gene replacement is decreased. All of the recombinants in the rad52-8 strains that would be categorized as gene replacements based on their phenotype are diploids apparently derived by endomitosis and excisive recombination. Studies on rad1 delta rad52-8 strains show that these mutations interact synergistically in the presence or absence of HOT1, resulting in low levels of recombination. The rate of gene replacement but not excisive recombination is stimulated by HOT1 in rad1 delta rad52-8 strains. Taken together, the results show that HOT1 stimulates exchange using multiple recombination pathways. Some of the activity of HOT1 is RAD1-dependent, some is RAD52-dependent, and some requires either RAD1 or RAD52 as suggested by the synergistic interaction found in double mutant strains. There is also a component of HOT1 activity that is independent of both RAD1 and RAD52.


Subject(s)
RNA Polymerase I/metabolism , Recombination, Genetic , Regulatory Sequences, Nucleic Acid , Saccharomyces cerevisiae/genetics , Blotting, Southern , Diploidy , Genes, Fungal , Mutation , Phenotype , RNA Polymerase I/genetics , Saccharomyces cerevisiae/enzymology
13.
Genetics ; 124(2): 263-73, 1990 Feb.
Article in English | MEDLINE | ID: mdl-2407612

ABSTRACT

Many genetic studies have shown that the frequency of homologous recombination depends largely on the distance in which recombination can occur. We have studied the effect of varying the length of duplicated sequences on the frequency of mitotic intrachromosomal recombination in Saccharomyces cerevisiae. We find that the frequency of recombination resulting in the loss of one of the repeats and the intervening sequences reaches a plateau when the repeats are short. In addition, the frequency of recombination to correct a point mutation contained in one of these repeats is not proportional to the size of the duplication but rather depends dramatically on the location of the mutation within the repeated sequences. However, the frequency of mitotic interchromosomal reciprocal recombination is dependent on the distance separating the markers. The difference in the response of intrachromosomal and interchromosomal mitotic recombination to increasing lengths of homology may indicate there are different rate-limiting steps for recombination in these two cases. These findings have important implications for the maintenance and evolution of duplicated sequences.


Subject(s)
Chromosomes, Fungal , Mitosis , Recombination, Genetic , Saccharomyces cerevisiae/genetics , Chromosomes, Fungal/ultrastructure , DNA, Fungal/genetics , Mutation , Repetitive Sequences, Nucleic Acid , Restriction Mapping , Transformation, Genetic
14.
Cell ; 48(6): 1071-9, 1987 Mar 27.
Article in English | MEDLINE | ID: mdl-3548996

ABSTRACT

A DNA sequence (HOT1) from the repeated ribosomal RNA gene cluster of Saccharomyces cerevisiae can stimulate genetic exchange when inserted at novel locations in the yeast genome. Localization of the sequences required for HOT1 activity demonstrates that two noncontiguous fragments of DNA are required for the stimulation of recombination. One of these fragments contains the transcription initiation site for the major 35S ribosomal RNA precursor. The other contains an enhancer of RNA polymerase I transcription. We suggest that transcription by RNA polymerase I initiating in the inserted rDNA and proceeding through the adjacent sequences is responsible for the stimulation of exchange. Consistent with this interpretation, insertion of the putative termination site for RNA polymerase I transcription between HOT1 and the adjacent recombining DNA abolishes the recombination stimulation. Transcription through both copies of the homologous recombining sequences appears to be necessary for enhanced exchange.


Subject(s)
DNA, Ribosomal/genetics , Genes, Fungal , Genes, Regulator , RNA Polymerase I/metabolism , Recombination, Genetic , Saccharomyces cerevisiae/genetics , Transcription, Genetic , Base Sequence , Plasmids
15.
Cell ; 39(2 Pt 1): 377-86, 1984 Dec.
Article in English | MEDLINE | ID: mdl-6094015

ABSTRACT

Special mechanisms for stimulating recombination among the nearly identical repeat units of certain multigene families may exist in order to maintain their sequence homogeneity. We have found evidence for such a recombination-stimulating activity in the tandemly repeated ribosomal RNA genes of yeast. A fragment of the yeast ribosomal DNA (rDNA), containing the 5S gene, nontranscribed spacer DNA, and part of the 25S gene, causes a localized stimulation of recombination when inserted at novel locations in the yeast genome. The rDNA fragment stimulates both interchromosomal and intrachromosomal mitotic recombination but not meiotic recombination. To stimulate mitotic recombination, the fragment must act on both copies of the recombining gene. Furthermore, the rDNA fragment stimulates exchange only when inserted with the 5S gene proximal to, and the 25S gene distal to, the recombining alleles.


Subject(s)
DNA, Ribosomal/genetics , Genes, Fungal , Recombination, Genetic , Saccharomyces cerevisiae/genetics , Base Sequence , Chromosomes/physiology , DNA Restriction Enzymes , DNA Transposable Elements , Plasmids
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