Biopolym. Cell. 1991; 7(3):13-20.
RecA-like proteins in nuclei of higher eukaryotes: detection of pachytene chromosome cores in isolated unfixed bovine meiotic nuclei with antibodies to RecA protein of Escherichia coli
1Engelhardt P., 2Akhmedov A. T., 2Lanzov V. A.
  1. University of Helsinki
    Helsinki, Finland
  2. B. P. Konstantinov Institute of Nuclear Physics, Academy of Sciences of the USSR
    Gatchina, Leningrad distr., USSR

Abstract

Polyvalent antibodies to RecA protein of E. coli (anti-RecA) interect specifically with 40-and 90-kDa polypeptides from pachylene nuclei of bull testes. As it was shown by the indirect immunofluorescence technique, using of the anti-RecA in unfixed isolated nuclei at different mciotic stags revealed the uniform staining of fin ineiotic chromosome cores with bright staining dots (sometimes of telomeric localization) surrounded by DNA counterstaing with propidium iodide. This picture suggests binding of anii-RecA with components of synaptonemal complexes that is expressed more brightly in the recombination nodules during chiasma formation. The staining by anti-RecA was more feeble and difiusible in premeiotic cells. In spermatozoa antigenes are revealed in the dorsal or acrosome-like material of their heads.

References

[1] Cox MM, Lehman IR. Enzymes of general recombination. Annu Rev Biochem. 1987;56:229-62.
[2] Kenne K, Ljungquist S. A DNA-recombinogenic activity in human cells. Nucleic Acids Res. 1984;12(7):3057-68.
[3] Hotta Y, Tabata S, Bouchard RA, Piñon R, Stern H. General recombination mechanisms in extracts of meiotic cells. Chromosoma. 1985;93(2):140-51.
[4] Cassuto E, Lightfoot LA, Howard-Flanders P. Partial purification of an activity from human cells that promotes homologous pairing and the formation of heteroduplex DNA in the presence of ATP. Mol Gen Genet. 1987;208(1-2):10-4.
[5] Ganea D, Moore P, Chekuri L, Kucherlapati R. Characterization of an ATP-dependent DNA strand transferase from human cells. Mol Cell Biol. 1987;7(9):3124-30.
[6] Hsieh P, Meyn MS, Camerini-Otero RD. Partial purification and characterization of a recombinase from human cells. Cell. 1986;44(6):885-94.
[7] Lopez B, Rousset S, Coppey J. Homologous recombination intermediates between two duplex DNA catalysed by human cell extracts. Nucleic Acids Res. 1987;15(14):5643-55.
[8] Moore SP, Rich A, Fishel R. The human recombination strand exchange process. Genome. 1989;31(1):45-52.
[9] Akhmedov AT, Namsaraev EA, Zaitseva EM, Zaiisev EN, Lantsov VA. Study of recombination activity in mammalian cell extracts. Biopolym Cell. 1990; 6(2):38-45.
[10] Higashitani A, Tabata S, Ogawa T, Ogawa H, Shibata M, Hotta Y. ATP-independent strand transfer protein from murine spermatocytes, spermatids, and spermatozoa. Exp Cell Res. 1990;186(2):317-23.
[11] Giroux CN. Chromosome synapsis and meiotic recombination. Genetic recombination. Eds R. Kucherlapati, G. Smith. Washington: Amer. Soc. Microbiol., 1988: 465-96.
[12] Angulo JF, Moreau PL, Maunoury R, Laporte J, Hill AM, Bertolotti R, Devoret R. KIN, a mammalian nuclear protein immunologically related to E. coli RecA protein. Mutat Res. 1989;217(2):123-34.
[13] Hotta Y, Stern H. Meiotic protein in spermatocytes of mammals. Nat New Biol. 1971;234(46):83-6.
[14] Cox MM, McEntee K, Lehman IR. A simple and rapid procedure for the large scale purification of the recA protein of Escherichia coli. J Biol Chem. 1981;256(9):4676-8.
[15] Jaton JC, Brandt DCh., Vassalli P. The isolation and character- ization of immunoglobulins, antibodies, and their constituent polypeptide chains, in Immunological Methods, vol. 1 (Lefkovits, I. and Pernis, B., eds.), Academic, New York, 1979:43-67.
[16] ZaÄ­tsev EN, ZaÄ­tseva EM, Bakhlanova IV, Gorelov VN, Kuz'min NP. Cloning and characteristics of recA gene in Pseudomonas aeruginosa. Genetika. 1986;22(11):2721-7.
[17] Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-5.
[18] Engelhardt P, Plagens U, Zbarsky IB, Filatova LS. Granules 25-30 nm in diameter: basic constituent of the nuclear matrix, chromosome scaffold, and nuclear envelope. Proc Natl Acad Sci U S A. 1982;79(22):6937-40.
[19] Thelander L, Berg P. Isolation and characterization of expressible cDNA clones encoding the M1 and M2 subunits of mouse ribonucleotide reductase. Mol Cell Biol. 1986;6(10):3433-42.
[20] Angulo JF, Schwencke J, Moreau PL, Moustacchi E, Devoret R. A yeast protein analogous to Escherichia coli RecA protein whose cellular level is enhanced after UV irradiation. Mol Gen Genet. 1985;201(1):20-4.
[21] Elledge SJ, Davis RW. Identification and isolation of the gene encoding the small subunit of ribonucleotide reductase from Saccharomyces cerevisiae: DNA damage-inducible gene required for mitotic viability. Mol Cell Biol. 1987;7(8):2783-93.
[22] Engström Y, Rozell B, Hansson HA, Stemme S, Thelander L. Localization of ribonucleotide reductase in mammalian cells. EMBO J. 1984;3(4):863-7. PubMed
[23] Heyting C, Moens PB, van Raamsdonk W, Dietrich AJ, Vink AC, Redeker EJ. Identification of two major components of the lateral elements of synaptonemal complexes of the rat. Eur J Cell Biol. 1987;43(1):148-54.
[24] Heyting C, Dettmers RJ, Dietrich AJ, Redeker EJ, Vink AC. Two major components of synaptonemal complexes are specific for meiotic prophase nuclei. Chromosoma. 1988;96(4):325-32.
[25] Moens PB, Heyting C, Dietrich AJ, van Raamsdonk W, Chen Q. Synaptonemal complex antigen location and conservation. J Cell Biol. 1987;105(1):93-103.
[26] Moens PB, Earnshaw WC. Anti-topoisomerase II recognizes meiotic chromosome cores. Chromosoma. 1989;98(5):317-22.