Abstract
The chromatin fraction of transcriptionally active macronuclei ofStylonychia lemnae shows a typical core histone pattern after SDS-polyacrylamide gel electrophoresis (SDS-PAGE). However, extracts of transcriptionally inactive micronuclei display only two bands in the molecular weight range of macronuclear core histones, corresponding to macronuclear H2A/H2B and H4. While a band corresponding to macronuclear H3 cannot be detected, the micronuclear fraction contains a prominent Mr 21,000 protein (protein X). We examined the chromatin fractions of the two kinds of nuclei in detail using specific antibodies directed against core histones and synthetic peptides of histones. Both microand macronuclei contain histones that are specifically recognized by antibodies directed against H2A, H2B and H4. Antigenicity in association with core histone H3 is found only in the macronuclear anlagen and in macronuclear extracts. None of the antibodies used react with the micronucleus specific protein X. During macronuclear development protein X disappears while macronuclear H3 appears. Thus we speculate, that protein X may be functionally related to macronuclear H3. The inactive state of the micronucleus is also characterized by the presence of two proteins, Y1 and Y2, with apparent Mrs of 45,000 in SDS-polyacrylamide gels. These proteins are recognized by antibodies raised against the N-terminal peptide of H3 and the C-terminal peptide of H2B respectively. Y1 and Y2 are also present during all of the transcriptionally inactive stages of macronuclear development but are not found in the active macronucleus. By these means chromatin extracts of micronuclei, macronuclear anlagen and macronuclei can be clearly distinguished. The observed differences reveal a sequence of changes in chromatin organization that correlate with the transition from the transcriptionally inactive to the transcriptionally active state of the chromatin.
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References
Allis CD, Wiggins JC (1984) Histone rearrangements accompany nuclear differentiation and dedifferentiation inTetrahymena. Dev Biol 101:282–294
Allis CD, Glover CVC, Gorovsky MA (1979) Micronuclei ofTetrahymena contain two types of histone H3. Proc Natl Acad Sci USA 76:4857–4861
Allis CD, Wiggins JC, Chicoine LG, Richman R (1984) Proteolytic processing of H1-like histones in chromatin: a physiologically and developmentally regulated event inTetrahymena micronuclei. J Cell Biol 99:1669–1677
Ammermann D, Schlegel M (1983) Characterization of two sibling species of the genusStylonychia (Ciliata, Hypotricha):S. mytilus Ehrenberg 1838 andS. lemnae n. sp. I. Morphology and reproductive behavior. J Protozool 30:290–294
Ammermann D, Steinbrück G, Berger L v, Hennig W (1974) The development of the macronucleus in the ciliated protozoanStylonychia mytilus. Chromosoma 45:401–429
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding. Anal Biochem 72:248–254
Butler AP, Laughlin TJ, Cadilla CL, Henry JM, Olins DE (1984) Physical structure of gene-sized chromatin from the protozoanOxytricha. Nucleic Acids Res 12:3201–3217
Cadilla CL, Harp J, Flanagan JM, Olins AL, Olins DE (1986) Preparation and characterization of soluble macronuclear chromatin from the hypotrichEuplotes eurystomus. Nucleic Acids Res 14:823–841
Caplan E (1975) A very rapidly migrating f1 histone associated with gene sized pieces of DNA in the macronucleus ofOxytricha sp. Biochim Biophys Acta 407:109–113
Caplan E (1977) Histones and other basic nuclear proteins in genetically active and inactive nuclei to the ciliateOxytricha sp. Biochim Biophys Acta 479:214–219
Doenecke D (1988) Histones, histone variants and postsynthetic histone modifications. In: Kahl G (ed) Architecture of eukaryotic genes. VCH Verlagsgemeinschaft, Weinheim, pp 123–141
Gorovsky MA (1986) Ciliate chromatin and histones. In: Gall J (ed) The molecular biology of ciliated protozoa. Academic Press, Orlando, pp 227–256
Gorovsky MA, Keevert JB, Pleger GL (1974) Subunit structure of a naturally occuring chromatin lacking histones F1 and F3. J Cell Biol 61:134–145
Hermann AL, Cadilla CL, Cacheiro LH, Carne AF, Olins DE (1987) An H1-like protein from the macronucleus ofEuplotes eurystomus. Eur J Cell Biol 43:155–162
Kraut H, Lipps HJ (1984) Macronucleus reorganization during sexual reproduction in hypotrichous ciliates. In: Engels W (ed) Advances in invertebrate reproduction, vol 3. Elsevier, Amsterdam, pp 533–540
Klobutcher LA, Prescott DM (1986) The special case of the hypotrichs. In: Gall J (ed) The molecular biology of ciliated protozoa. Academic Press, Orlando, pp 111–154
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lipps HJ, Handtke KG (1975) Studies on the histones of the ciliateStylonychia mytilus. Chromosoma 49:303–320
Lipps HJ, Morris NR (1977) Chromatin structure in the nuclei of of the ciliateStylonychia mytilus. Biochem Biophys Res Commun 74:230–234
Lipps HJ, Sapra GR, Ammermann D (1974) The histones of the ciliated protozoanStylonychia mytilus. Chromosoma 45:273–280
Meyer GF, Lipps HJ (1980) Chromatin elimination in the hypotrichous ciliate,Stylonychia mytilus. Chromosoma 77:285–297
Meyer GF, Lipps HJ (1981) The formation of polytene chromosomes during macronucleus development of the hypotrichous ciliate,Stylonychia mytilus. Chromosoma 82:309–314
Muller S (1988) Immunization with peptides. In: Burdon RH, Van Knippenberg PM (eds) Laboratory techniques in biochemistry and molecular biology, vol 19. Elsevier, Amsterdam, pp 131–144
Muller S, Van Regenmortel MHV (1989) Use of antihistone antibodies with nucleosomes. Methods Enzymol 170:251–263
Muller S, Himmelspach K, Van Regenmortel MHV (1982a) Immunochemical localization of the C-terminal hexapeptide of histone H3 at the surface of chromatin subunits. EMBO J 1:421–425
Muller S, Erard M, Burggraf E, Couppez M, Sautiere P, Champagne M, Van Regenmortel MHV (1982b) Immunochemical detection of changes in chromatin subunits induced by histone H4 acetylation. EMBO J 1:939–944
Muller S, Mazen A, Martinage A, Van Regenmortel MHV (1984) Use of histone antibodies for studying chromatin topography and the phosphorylation of chromatin subunits. EMBO J 3:2431–2436
Muller S, Couppez M, Briand JP, Gordon J, Sautiere P, Van Regenmortel MHV (1985) Antigenic structure of histone H2B. Biochim Biophys Acta 827:235–246
Muller S, Plaué S, Couppez M, Van Regenmortel MHV (1986) Comparison of different methods for localizing antigenic regions in histone H2A. Mol Immunol 23:593–601
Muller S, Isabey A, Couppez M, Plaué S, Sommermeyer G, Van Regenmortel MHV (1987) Specifity of antibodies raised against triacetylated histone H4. Mol Immunol 24:779–789
Ruder FJ, Frasch M, Mettenleiter TC, Büsen W (1987) Appearance of two maternally directed histone H2A variants precedes zygotic ubiquitination of H2A in early embryogenesis ofSciara coprophila (Diptera). Dev Biol 122:568–576
Steinbrück G (1986) Molecular reorganization during nuclear differentiation in ciliates. In: Hennig W (ed) Results and problems in cell differentiation, vol 13. Springer, Berlin, Heidelberg New York, pp 105–174
White E, Shapiro DL, Allis CD, Gorovsky MA (1988) Sequence and properties of the message encodingTetrahymena hv1, a highly evolutionary conserved histone H2A variant that is associated with active genes. Nucleic Acids Res 16:179–197
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Schlegel, M., Muller, S., Ruder, F. et al. Transcriptionally inactive micronuclei, macronuclear anlagen and transcriptionally active macronuclei differ in histone composition in the hypotrichous ciliateStylonychia lemnae . Chromosoma 99, 401–406 (1990). https://doi.org/10.1007/BF01726691
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DOI: https://doi.org/10.1007/BF01726691