Homologous recombination in fission yeast: Absence of crossover interference and synaptonemal complex

The study of homologous recombination in the fission yeastSchizosaccharomyces pombe has recently been extended to the cytological analysis of meiotic prophase. Unlike in most eukaryotes no tripartite SC structure is detectable, but linear elements resembling axial cores of other eukaryotes are retained. They may be indispensable for meiotic recombination and proper chromosome segregation in meiosis I. In addition fission yeast shows interesting features of chromosome organization in vegetative and meiotic cells: Centromeres and telomeres cluster and associate with the spindle pole body. The special properties of fission yeast meiosis correlate with the absence of crossover interference in meiotic recombination. These findings are discussed. In addition homologous recombination in fission yeast is reviewed briefly.This article is dedicated to Urs Leupold, the founder of fission yeast genetics.     

Progressive resistance against the male recombination factor 31.1 MRF acquired by Drosophila melanogaster.

A genetically abnormal but structurally normal second chromosome (31.1 MRF) which induces recombination in the male sex, was introduced by outcrossing into the cytoplasm of a normal strain. In this new combination, male recombination frequency induced by females dropped from 3.81% to 0.17% within 10 generations, indicating a gradual acquisition of resistance against the activities of the 31.1 MRF.     

Telomeres and DNA double-strand breaks: ever the twain shall meet?

Telomeres were first recognized as a bona fide constituent of the chromosome based on their inability to rejoin with broken chromosome ends produced by radiation. Today, we recognize two essential and interrelated properties of telomeres. They circumvent the so-called end-replication problem faced by genomes composed of linear chromosomes, which erode from their termini with each successive cell division. Equally vital is the end-capping function that telomeres provide, which is necessary to deter chromosome ends from illicit recombination. This latter property is critical in facilitating the distinction between the naturally occurring DNA double-strand breaks (DSBs) found at chromosome ends (i.e., telomeres) and DSBs produced by exogenous agents. Here we discuss, in a brief historical narrative, key discoveries that led investigators to appreciate the unique properties of telomeres in protecting chromosome ends, and the consequences of telomere dysfunction, particularly as related to recombination involving radiation-induced DSBs. Dedication. In appreciation of his heart-felt commitment to research and education, and the life-long influence he has had on the lives of students and colleagues, the authors wish to dedicate this paper to Professor Joel S. Bedford. Received 21 May 2007; received after revision 28 June 2007; accepted 6 August 2007     

Chromosomal localization of the locus PGM (phosphoglucomutase) inDrosophila buzzatii

Summary The locus PGM ofD. buzzatii is localized in the linkage group of chromosome 4, outside the region blocked by the inversion 4s and with a recombination percentage of about 16% from the inversion breakpoint.     

The search for the right partner: Homologous pairing and DNA strand exchange proteins in eukaryotes

Finding the right partner is a central problem in homologous recombination. Common to all models for general recombination is a homologous pairing and DNA strand exchange step. In prokaryotes this process has mainly been studied with the RecA protein ofEscherichia coli. Two approaches have been used to find homologous pairing and DNA strand exchange proteins in eukaryotes. A biochemical approach has resulted in numerous proteins from various organisms. Almost all of these proteins are biochemically fundamentally different from RecA. The in vivo role of these proteins is largely not understood. A molecular-genetical approach has identified structural homologs to theE. coli RecA protein in the yeastSaccharomyces cerevisiae and subsequently in other organisms including other fungi, mammals, birds, and plants. The biochemistry of the eukaryotic RecA homologs is largely unsolved. For the fungal RecA homologs (S. cerevisiae RAD51, RAD55, RAD57, DMC1; Schizosaccharomyces pombe rad51; Neurospora crassa mei3) a role in homologous recombination and recombinational repair is evident. Besides recombination, homologous pairing proteins might be involved in other cellular processes like chromosome pairing or gene inactivation.     

Recombination frequency and DNA content of the distal part of the second chromosome ofDrosophila hydei sturtevant

Summary Four visible markers, including a newly isolated one, have been cytologically mapped on the second chromosome ofDrosophila hydei. Both the frequency of recombination and the amount of DNA between these markers have been determined. From these data the coefficient of exchange has been calculated.     

Homologous recombination in plants

In plants three different approaches have been used to study homologous DNA recombination; extrachromosomal recombination (ECR) between transfected DNA molecules, intrachromosomal recombination (ICR) between repeated genes integrated into and resident at the genome and recombination between introduced DNA and homologous sequences in the genome (gene targeting). ECR is efficient (10^–1 to 10^–3) and occurs mainly during a limited time period early after transfection. It proceeds predominantly via nonconservative single-strand annealing. ICR, which in most cases is described best by the double-strand break repair model of recombination, occurs at frequencies of one event in 10⁵ to 10⁷ cells. ICR takes place throughout the whole life-cycle of a plant, in all organs and at different developmental stages. As there exists no predetermined germline in plants, somatic recombination events can be transferred to the next generation. Recombination frequencies are enhanced by DNA damage. Gene targeting, like ICR, occurs at low rates in plant cells. Almost nothing is known about the enzymes involved in homologous recombination in plants.     

Chromosome 3-linked frontotemporal dementia

Frontotemporal dementia accounts for a significant minority of all cases of presenile dementia. Many pedigrees have been described in which frontotemporal dementia is inherited as an autosomal dominant trait. Frontotemporal dementia is genetically heterogeneous with loci identified on chromosome 17 and chromosome 3. Clinical, pathological and genetic findings are described in a large Danish family in which the disease gene lies in the pericentromeric region of chromosome 3.     

Genetic characterization of the new morphological and UV-sensitive mutants in Coprinus cinereus. I. A UV-sensitive mutation rad 1 associated with elevated frequencies of mitotic and meiotic recombination

Studies on the effect of an UV-sensitive mutation, rad 1, in meiotic and mitotic recombination in Coprinus indicated that, in homozygous condition, rad 1 increased the spontaneous meiotic recombination by 50% and UV-induced mitotic intergenic recombination by about 5-fold. The homozygous rad 1 diploid was shown to be much more sensitive to the recombinogenic effects of polyfunctional than than of mono- or non-functional alkylating agents.     

Genetic analysis of interspecific crosses Mus musculus L. x Mus spretus Lataste: linkage of Adh-1 with Amy-1 on chromosome 3 and Es-14 with Mod-1 on chromosome 9]

192 offsprings from interspecific back-crosses (male M. spretus x female BALB/c) F1 x male BALB/c or (male M. spretus x female C57BL6) F1 x male C57BL6 were analysed at thirteen structural loci. Linkage of ES-14 with Mod-1 on chromosome 9 and that of Adh-1 with Amy-1 on chromosome 3 are shown. The following order centromere/Car-2/Amy-1 is tentatively proposed for these loci on chromosome 3.     

Hotspots of homologous recombination

Homologous recombination occurs at higher than average frequency at and near hotspots. Hotspots are special nucleotide sequences recognized by proteins that promote, directly or indirectly, a rate limiting step of recombination. This review focuses on two well-studied examples, the Chi sites of the bacteriumEscherichia coli and the M26 site of the fission yeastSchizosaccharomyces pombe. Chi, 5 G-C-T-G-G-T-G-G 3, is recognized by the RecBCD enzyme, which nicks the DNA near Chi and produces a 3-ended single-stranded DNA tail; this tail is a potent substrate for homologous pairing by RecA and single-stranded DNA binding proteins. M26, 5 A-T-G-A-C-G-T 3, is recognized by a heterodimeric protein and stimulates, by an as-yet-unknown mechanism, meiotic recombination at and near theade6 gene. Additional hotspots in bacteria, fungi, and mammals enhance recombination directly or indirectly via a variety of mechanisms. Although hotspots are widespread among organisms, the biological role of their localized enhancement of recombination remains a matter of speculation.     

A further study of LDH isozymes in theRana esculenta complex

Summary Electrophoretic analyses of LDH isozymes in the 3 types of European green frogs by using the buffer system of Williams and Reisfeld demonstrated the occurrence of 4 allelic genes coding for the B subunits. Based on the distribution of these alleles and results from dissociation and recombination of the subunits, the hybrid nature of Rana esculenta is confirmed.This study was supported in part by the Swiss National Science Foundation and the Georges-und-Antoine-Claraz-Schenkung.     

Epithelial-mesenchymal interaction in differentiation of duodenal epithelium of fetal rats in organ culture

Summary Epithelial-mesenchymal interaction in the differentiation of duodenal epithelium of fetal rats was investigated by recombination experiments in vitro. The proportion of goblet cells in duodenal epithelium was significantly greater on recombination of developing duodenal epithelium with mesenchyme of the glandular stomach than on recombination with that of the duodenum. Mesenchyme of the glandular stomach or forestomach was better than duodenal mesenchyme in supporting morphogenesis of duodenal epithelium. Treatment of tissues with N-methyl-N-nitro-N-nitrosoguanidine (MNNG) did not affect these tissue interactions.This work was supported by grants-in-aid for cancer research from the Ministry of Education, Science and Culture, Japan.Acknowledgment. The authors wish to express their gratitude to Prof. T. Mizuno of the University of Tokyo for valuable suggestions.     

Induction of recombination between diverged sequences in a mammalian genome by a double-strand break

To investigate whether mammalian cells can carry out recombinational double-strand break (DSB) repair between highly diverged sequences, mouse fibroblasts were transfected with DNA substrates that contained a “recipient” thymidine kinase (tk) gene disrupted by the recognition site for endonuclease I-SceI. Substrates also contained a linked “donor” tk gene sequence. Following DSB induction by I-SceI, selection for tk-expressing clones allowed recovery of repair events occurring by nonhomologous end-joining or recombination with the donor sequence. Although recombinational repair was most efficient when donor and recipient shared near-perfect homology, we recovered recombination events between recipient and donor sequences displaying 20 % nucleotide mismatch. Recombination between such imperfectly matched (“homeologous”) sequences occurred at a frequency of 1.7 × 10^?7 events per cell and constituted 3 % of the DSB repair events recovered with the pair of homeologous sequences. Additional experiments were done with a substrate containing a donor sequence comprised of a region sharing high homology with the recipient and an adjacent region homeologous to the recipient. Recombinational DSB repair tracts initiating within high homology propagated into homeology in 11 of 112 repair events. These collective results contrasted with our earlier work in which spontaneous recombination (not intentionally induced by a DSB) between homeologous sequences occurred at an undetectable frequency of less than 10^?9 events per cell, and in which events initiating within high homology propagated into adjoining homeology in one of 81 events examined. Our current work suggests that homology requirements for recombination are effectively relaxed in proximity to a DSB in a mammalian genome.     

New inversion of the pair no. 3 chromosome in a black rat

Summary A Japanese black rat (Rattus rattus tanezumi) with a subtelocentric pair no. 3 chromosome was found in Gotenba, Japan. By comparison of the length in both members of the chromosome pair, and from the G-ban pattern, the subtelocentrics seemed to have developed from the original acrocentrics by the pericentric inversion.The author wishes to express his sincere thanks to Dr. K. Moriwaki for the help of the collection of the material and also for the skilful technical help of Miss Y. Ochiai. Supported by a grant-in-aid from the Ministry of Education, Science and Culture of Japan (Nos 111510 and 111506). Contribution No. 1125 from the National Institute of Genetics, Japan.     

Accumulation of drastic mutants in selection lines for resistance to the insecticides dichlorvos and malathion inDrosophila melanogaster

Summary Viability tests were performed on second and third chromosomes from lines ofDrosophila melanogaster selected for increased resistance to the organophosphorus insecticides dichlorvos and malathion, in order to evaluate the accumulation of drastic alleles. Our results show that malathion reduces significantly the relative viability of chromosome 3 and also increases the frequency of drastic alleles in this chromosome, while dichlorvos increases significnatly the frequency of drastic alleles in chromosome 2.Work supported in part by the Spanish Ministry of Education and Science (Grant No. 0577/84 CAICYT).—We are grateful to Productos Cruz Verde S.A. and Agrocrós S.A. for generously supplying the insecticides dichlorvos and malathion, respectively.     

Genetic characterization of the new morphological and UV-sensitive mutants inCoprinus cinereus. I. A UV-sensitive mutationrad1 associated with elevated frequencies of mitotic and meiotic recombination

Summary Studies on the effect of an UV-sensitive mutation,rad1, in meiotic and mitotic recombination inCoprinus indicated that, in homozygous condition,rad1 increased the spontaneous meiotic recombination by 50% and UV-induced mitotic intergenic recombination by about 5-fold. The homozygousrad1 diploid was shwon to be much more sensitive to the recombinogenic effects of polyfunctional than of mono- or non-functional alkylating agents.This research, carried out in partial fulfilment of Ph.D. requirements at the University of London, was supported by a grant from The School of Public Health, University of Tehran, Iran. My thanks are due to Dr J.W. Cowan for his advice and constructive criticism. I am also indebted to Dr A.B. Borkovec of The Institute of Chemosterilants Laboratory, US Ministry of Agriculture for the most generous supply of aziridinyl phosphine compounds.     

Zur Frage der Wirkung von DTPA auf Säugerchromosomen in vitro

Summary The influence of DTPA on the chromosome aberration rate of Chinese hamster cells in culture was studied. No increase of the aberration rate was observed after treatment with 10^–2 and 10^–3 M concentrations.