Meiotic synapsis in the absence of recombination

Science

Volumn 279, Issue 5352, 1998, Pages 876-878

  McKim, Kim S ^a   Green Marroquin, Becky L ^a   Sekelsky, Jeff J ^a   Chin, Gregory ^a   Steinberg, Carrie ^a   Khodosh, Rita ^a   Scott Hawley, R ^a  

^a UNIVERSITY OF CALIFORNIA   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

DOUBLE STRANDED DNA;

ANIMAL CELL; ARTICLE; CHROMOSOME PAIRING; CONTROLLED STUDY; CROSSING OVER; DNA STRAND BREAKAGE; DROSOPHILA MELANOGASTER; FEMALE; GENE CONVERSION; GENETIC RECOMBINATION; MEIOSIS; NONHUMAN; OOCYTE; PRIORITY JOURNAL; SACCHAROMYCES CEREVISIAE; SYNAPTONEMAL COMPLEX; TRANSMISSION ELECTRON MICROSCOPY;

ANIMALS; CHROMOSOMES; CROSSING OVER, GENETIC; DROSOPHILA MELANOGASTER; FEMALE; GENE CONVERSION; MEIOSIS; MUTATION; OOCYTES; RECOMBINATION, GENETIC; SACCHAROMYCES CEREVISIAE; SISTER CHROMATID EXCHANGE; SYNAPTONEMAL COMPLEX;

ANIMALIA; DROSOPHILA MELANOGASTER; MELANOGASTER; SACCHAROMYCES CEREVISIAE;

EID: 0032488883     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.279.5352.876     Document Type: Article

References (49)

1. R. S. Hawley and T. Arbel, Cell 72, 301 (1993).
2. G. S. Roeder, Proc. Natl. Acad. Sci. U.S.A 92, 10450 (1995).
3. N. Kleckner, ibid. 93, 8167 (1996).
4. R. Padmore, L. Cao, N. Kleckner, Cell 66, 1239 (1991).
5. B. Rockmill and G. S. Roeder, Genetics 126, 563 (1990); M. Sym and G. S. Roeder, Cell 79, 283 (1994).
6. B. Rockmill and G. S. Roeder, Genetics 126, 563 (1990); M. Sym and G. S. Roeder, Cell 79, 283 (1994).
7. B. de Massy, F. Baudat, A. Nicolas, Proc. Natl. Acad. Sci. U.S.A. 91, 11929 (1994); L. A. Gilbertson and F. W. Stahl, ibid., p. 11934.
8. B. de Massy, F. Baudat, A. Nicolas, Proc. Natl. Acad. Sci. U.S.A. 91, 11929 (1994); L. A. Gilbertson and F. W. Stahl, ibid., p. 11934.
9. S. Jinks-Robertson and T. D. Petes, Genetics 114, 731 (1986); M. Lichten, R. H. Borts, J. E. Haber, ibid. 115, 233 (1987).
10. S. Jinks-Robertson and T. D. Petes, Genetics 114, 731 (1986); M. Lichten, R. H. Borts, J. E. Haber, ibid. 115, 233 (1987).
11. The isolation and initial characterization of the second chromosome recessive mutation, mei-W68, was performed by B. S. Baker (26). The allele studied here is the more severe of the two existing alleles. The third chromosomal recessive mutant, mei-P22, was isolated in a large scale P-element mutagenesis screen for mutants with a high frequency of X-chromosome nondisjunction in the female germline (27). Subsequent analysis has shown that this mutant is caused by the insertion of a P-element (28).
12. Because of the low frequency of meiotic recombination in small intervals, gene conversion events in D. melanogaster are rare. The rosy system facilitates the isolation of intragenic recombinants, including gene conversions, because of a selection system for wild-type recombinants (29). Females with the genotypes shown in Table 1 were crossed to either TM2, ry/MKRS, ry kar males as virgins, or as nonvirgins to their phenotypically rosy brothers. These crosses were done in bottles with 30 females and 30 males for the controls, or 50 females and 50 males for the meiotic mutants. The flies were transferred to new bottles every 3 days at which time 0.75 ml of 0.2% purine (Sigma) was added to the media. A total of five broods were set. To estimate the total number of progeny scored in each experiment, all the progeny were scored from the ∼5% of the bottles that were not given the purine treatment.
13. An exact estimate of the frequency of meiotic exchange in mei-W68 and mei-P22 mutants was complicated by the presence of premeiotic-exchange events. This problem is illustrated by an example with mei-P22. In our analysis of X-chromosome crossing over in females homozygous for mei-P22, 25 crossovers were observed among 1935 regular progeny. Twenty-three of the crossovers occurred in clusters (one cluster of 18 identical recombinants and one cluster of 5 identical recombinants). Similar results were obtained on the left arm of chromosome 2 and in two sets of data regarding the effect of the mei-W68 mutation on second chromosome crossing over. Those recombinants arising in clusters can be ascribed to premeiotic (that is mitotic) recombination events, and thus the frequency of meiotic exchange in mei-P22 and mei-W68 females is either zero or very close to zero. In support of this conclusion, in males, which are normally achiasmatic, the mei-W68 mutation caused an increase in the spontaneous cross-over frequency to an amount similar to that in mei-W68 females (30). Although the clusters of recombinants from mei-P22 females were larger than from mei-W68, the similarity of the mei-P22 and mei-W68 meiotic phenotypes (effects on gene conversion, and reductions in crossing over combined with increases in clustered events and their effects on SC formation) suggests that any differences between mei-P22 and mei-W68 in the production of crossover progeny are probably the result of differences in their effects on mitotic recombination.
14. Of the nine mei-W68 nuclei that were in the right developmental stage to have late RNs (between the onset of organelle passage through the ring canals and overt oocyte determination), no RNs were observed (16), despite the fact that with an average number of late nodules of 3.37 per nucleus (18), about 30 were expected. Similarly, no RNs were observed in three mei-P22 oocytes examined (76), although 10 were expected. No early RNs were observed in the four mei-W68 nuclei at the stage expected to display early RNs (that is, just before and just after the onset of organellar passage), when 11 would have been expected. The absence of RNs in mei-W68 and mei-P22 oocytes is concordant with the null-recombination phenotype of these mutants and is consistent with the hypothesis that neither mutant initiates recombination events.
15. 17 in a similar experiment. Using a different ring-X chromosome [R(1)2], Hall observed a Ring/Rod ratio of 0.755 (n = 7552) in controls and an elevated ratio of 0.894 (n = 5355) in c(3)G females (37). One reasonable explanation for these observations is that the reduced ring recovery observed in the two control experiments reflects the background frequency of meiotic sister-chromatid exchange, and that mei-P22 and c(3)G actually inhibit these sister-chromatid exchange events, as they do interhomolog events, and in doing so increase the transmissibility of the ring-X chromosome.
16. K. S. McKim, J. K. Jang, R. S. Hawley, unpublished results.
17. C. W. Hinton and J. C. Lucchesi, Genetics 45, 87 (1960).
18. Among a total of 709 progeny from mei-P22 homozygous females, 11.6% were nullo-X exceptions and 18.3% diplo-X exceptions. Among a total of 1024 progeny from mei-W68 homozygous females, 20.7% were nullo-X exceptions and 18.0% diplo-X exceptions.
19. A. T. C. Carpenter, personal communication.
20. _, Chromosoma 51, 157 (1975).
21. _, Genetics 92, 511 (1979); Chromosoma 75, 259 (1979); Ciba Found. Symp. 182 223 (1994).
22. _, Genetics 92, 511 (1979); Chromosoma 75, 259 (1979); Ciba Found. Symp. 182 223 (1994).
23. _, Genetics 92, 511 (1979); Chromosoma 75, 259 (1979); Ciba Found. Symp. 182 223 (1994).
24. P. A. Roberts, Genetics 65, 429 (1970); ibid. 71, 401 (1972); R. S. Hawley, ibid. 94, 625 (1980); R. E. Rosenbluth and D. L. Baillie, ibid. 99, 415 (1981); C. R. Burnham, J. T. Stout, W. H. Weinheimer, R. V. Kowles, R. L. Phillips, ibid. 71, 111 (1972); M. P. Maguire, J. Theor. Biol. 106, 605 (1984).
25. P. A. Roberts, Genetics 65, 429 (1970); ibid. 71, 401 (1972); R. S. Hawley, ibid. 94, 625 (1980); R. E. Rosenbluth and D. L. Baillie, ibid. 99, 415 (1981); C. R. Burnham, J. T. Stout, W. H. Weinheimer, R. V. Kowles, R. L. Phillips, ibid. 71, 111 (1972); M. P. Maguire, J. Theor. Biol. 106, 605 (1984).
26. P. A. Roberts, Genetics 65, 429 (1970); ibid. 71, 401 (1972); R. S. Hawley, ibid. 94, 625 (1980); R. E. Rosenbluth and D. L. Baillie, ibid. 99, 415 (1981); C. R. Burnham, J. T. Stout, W. H. Weinheimer, R. V. Kowles, R. L. Phillips, ibid. 71, 111 (1972); M. P. Maguire, J. Theor. Biol. 106, 605 (1984).
27. P. A. Roberts, Genetics 65, 429 (1970); ibid. 71, 401 (1972); R. S. Hawley, ibid. 94, 625 (1980); R. E. Rosenbluth and D. L. Baillie, ibid. 99, 415 (1981); C. R. Burnham, J. T. Stout, W. H. Weinheimer, R. V. Kowles, R. L. Phillips, ibid. 71, 111 (1972); M. P. Maguire, J. Theor. Biol. 106, 605 (1984).
28. P. A. Roberts, Genetics 65, 429 (1970); ibid. 71, 401 (1972); R. S. Hawley, ibid. 94, 625 (1980); R. E. Rosenbluth and D. L. Baillie, ibid. 99, 415 (1981); C. R. Burnham, J. T. Stout, W. H. Weinheimer, R. V. Kowles, R. L. Phillips, ibid. 71, 111 (1972); M. P. Maguire, J. Theor. Biol. 106, 605 (1984).
29. P. A. Roberts, Genetics 65, 429 (1970); ibid. 71, 401 (1972); R. S. Hawley, ibid. 94, 625 (1980); R. E. Rosenbluth and D. L. Baillie, ibid. 99, 415 (1981); C. R. Burnham, J. T. Stout, W. H. Weinheimer, R. V. Kowles, R. L. Phillips, ibid. 71, 111 (1972); M. P. Maguire, J. Theor. Biol. 106, 605 (1984).
30. F. Sherman, C. Helms, Genetics 88, 689 (1978); J. E. Haber, W.-Y. Leung, R. H. Borts, M. Lichten, Proc. Natl. Acad. Sci. U.S.A. 88, 1120 (1991); A. S. H. Goldman and M. Lichten, Genetics 144, 43 (1996).
31. F. Sherman, C. Helms, Genetics 88, 689 (1978); J. E. Haber, W.-Y. Leung, R. H. Borts, M. Lichten, Proc. Natl. Acad. Sci. U.S.A. 88, 1120 (1991); A. S. H. Goldman and M. Lichten, Genetics 144, 43 (1996).
32. F. Sherman, C. Helms, Genetics 88, 689 (1978); J. E. Haber, W.-Y. Leung, R. H. Borts, M. Lichten, Proc. Natl. Acad. Sci. U.S.A. 88, 1120 (1991); A. S. H. Goldman and M. Lichten, Genetics 144, 43 (1996).
33. R. Hipeau-Jacquotte, D. L. Brutlag, F. Brégégère, Mol. Gen. Genet. 220, 140 (1989).
34. L. Craymer, Genetics 99, 75 (1981); ibid. 108, 573 (1984).
35. L. Craymer, Genetics 99, 75 (1981); ibid. 108, 573 (1984).
36. A. M. Rose, D. L. Baillie, J. Curran, Mol. Gen. Genet. 195, 52 (1984); K. S. McKim, A. M. Howell, A. M. Rose, Genetics 120, 987 (1988); K. S. McKim, K. Peters, A. M. Rose, ibid. 134, 749 (1993).
37. A. M. Rose, D. L. Baillie, J. Curran, Mol. Gen. Genet. 195, 52 (1984); K. S. McKim, A. M. Howell, A. M. Rose, Genetics 120, 987 (1988); K. S. McKim, K. Peters, A. M. Rose, ibid. 134, 749 (1993).
38. A. M. Rose, D. L. Baillie, J. Curran, Mol. Gen. Genet. 195, 52 (1984); K. S. McKim, A. M. Howell, A. M. Rose, Genetics 120, 987 (1988); K. S. McKim, K. Peters, A. M. Rose, ibid. 134, 749 (1993).
39. P. B. Moens, J. A. M. Heddle, H. H. Q. Heng, Genome 40, 770 (1997).
40. N. Kleckner, R. Padmore, D. K. Bishop, Cold Spring Harbor Symp. Quant. Biol. 56, 729 (1991).
41. B. S. Baker, personal communication.
42. R. S. Hawley et al., unpublished results.
43. K. S. McKim, unpublished results.
44. A. J. Hilliker, S. Clark, A. Chovnick, Genetics 129, 779 (1991).
45. T. Lutken and B. S. Baker, Mutat. Res. 61, 221 (1979)
46. J. Hall, Drosophila Information Service 52, 142 (1977).
47. D. Curtis and W. Bender, Genetics 127, 739 (1991).
48. 20, dehydrated with ethanol to 70%, and en bloc stained with 2% uranyl acetate dissolved in 70% ethanol overnight. Ovaries were embedded in Spurr's resin instead of Epon-Araldite. Whole ovaries were flat-embedded with the ovarioles positioned so that the long axis of the germarium was parallel to the cutting plane. The germarium end of the block was serial sectioned with a thickness of 100 nm per section. The sections were collected on formvar, carbon-coated copper single-slot grids (1 mm by 2 mm). The sections were then poststained with uranyl acetate (in 50% ethanol) for 20 min and concentrated lead citrate for 30 s, with thorough rinsing after each staining. The sections were examined by electron microscopy (Philips EM-410) and photographed at low magnification. When the cells with four ring canals were located, they were photographed at a magnification of 14,000.
49. We are grateful to A. T. C. Carpenter for advice throughout all aspects of this study and for supplying the EM data for SC formation in mei-W68 mutants. We also thank B. S. Baker for data on the initial genetic characterization of mei-W68; J. Haber, N. Kleckner, and T. Petes for critical reading of the manuscript; and T. Arbel, W. Hurley, and R. French for assistance in the isolation of mei-P22. K.S.M. was supported by a fellowship from the Medical Research Council (Canada). Supported by a grant from the NIH to R.S.H.