Abstract
The specific activity and subcellular distribution of marker enzymes for the main subcellular components were analysed in homogenates of synchronized hepatoma cells (Morris 7288c), obtained by selective detachment at mitosis combined with a metaphase block with Colcemid. Markers for lysosomes, mitochondrial outer membrane, plasma membrane and cytosol are synthesized throughout the cycle at the same rate as the bulk of cellular protein. Larger variations are observed for a Golgi marker; after a decrease around mitosis, the specific activity of galactosyltransferase increases steadily from middle G1-phase on, and at the end of G2-phase it is nearly twice that observed at the beginning of G1-phase. Our results show that synthesis of cytochrome oxidase may occur preferentially in G2-phase. Large modifications of the density distribution of lysosomes are observed during the cell cycle; the median equilibrium density of lysosomal markers decreases in G1-phase, and some increase in soluble activity occurs at the same time. Reverse changes occur progressively during S- and G2-phases. At mitosis, Golgi galactosyltransferase shows a more dispersed distribution, and modifications in the density distribution of endoplasmic-reticulum NADPH–cytochrome c reductase are observed. The latter can be most easily explained by a detachment of ribosomes from endoplasmic-reticulum membranes. No significant modifications occur in mitochondrial and plasma-membrane markers.
Full text
PDF








Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Amar-Costesec A., Beaufay H., Wibo M., Thinès-Sempoux D., Feytmans E., Robbi M., Berthet J. Analytical study of microsomes and isolated subcellular membranes from rat liver. II. Preparation and composition of the microsomal fraction. J Cell Biol. 1974 Apr;61(1):201–212. doi: 10.1083/jcb.61.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barrett A. J. An improved color reagent for use in Barrett's assay of Cathepsin B. Anal Biochem. 1976 Nov;76(50):374–376. doi: 10.1016/0003-2697(76)90298-0. [DOI] [PubMed] [Google Scholar]
- Bartholeyns J., Quintart J., Baudhuin P. Inhibition of the discharge of endocytosed protein from phagosomes into lysosomes in hepatoma cells exposed to dimerized ribonuclease A. Biochem J. 1979 Feb 15;178(2):433–442. doi: 10.1042/bj1780433. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baudhuin P., Leroy-Houyet M. A., Quintart J., Berthet P. Application of cluster analysis for characterization of spatial distribution of particles by stereological methods. J Microsc. 1979 Jan;115(1):1–17. doi: 10.1111/j.1365-2818.1979.tb00148.x. [DOI] [PubMed] [Google Scholar]
- Berg T., Melbye B., Johnsen S. R., Prydz H. Activity of lysosomal enzymes in the various cell cycle phases of synchronized HeLa cells. Exp Cell Res. 1975 Aug;94(1):106–110. doi: 10.1016/0014-4827(75)90536-4. [DOI] [PubMed] [Google Scholar]
- Berlin R. D., Oliver J. M., Walter R. J. Surface functions during Mitosis I: phagocytosis, pinocytosis and mobility of surface-bound Con A. Cell. 1978 Oct;15(2):327–341. doi: 10.1016/0092-8674(78)90002-8. [DOI] [PubMed] [Google Scholar]
- Blobel G., Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. doi: 10.1083/jcb.67.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bosmann H. B., Bernacki R. J. Glycosidase activity. Glycosidase acitivity in L5178Y mouse leukemic cells and the activity of acid phosphatase, beta-galactosidase and beta-N-acetylgalactosaminidase and beta-N-acetylglucosaminidase in a synchronous L5178Y cell population. Exp Cell Res. 1970 Aug;61(2):379–386. doi: 10.1016/0014-4827(70)90461-1. [DOI] [PubMed] [Google Scholar]
- Bosmann H. B. Mitochondrial biochemical events in a synchronized mammalian cell population. J Biol Chem. 1971 Jun 25;246(12):3817–3823. [PubMed] [Google Scholar]
- Bowers W. E., de Duve C. Lysosomes in lymphoid tissue. II. Intracellular distribution of acid hydrolases. J Cell Biol. 1967 Feb;32(2):339–348. doi: 10.1083/jcb.32.2.339. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chatterjee S., Velicer L. F., Sweeley C. C. Glycosphingolipid glycosyl hydrolases and glycosidases of synchronized human KB cells. J Biol Chem. 1975 Jul 10;250(13):4972–4979. [PubMed] [Google Scholar]
- Churchill J. R., Studzinski G. P. Thymidine as synchronizing agent. 3. Persistence of cell cycle patterns of phosphatase activities and elevation of nuclease activity during inhibition of DNA synthesis. J Cell Physiol. 1970 Jun;75(3):297–303. doi: 10.1002/jcp.1040750306. [DOI] [PubMed] [Google Scholar]
- Criss W. E. Rat liver adenosine triphosphate: adenosine monophosphate phosphotransferase activity. II. Subcellular localization of adenylate kinase isozymes. J Biol Chem. 1970 Dec 10;245(23):6352–6356. [PubMed] [Google Scholar]
- DE DUVE C., PRESSMAN B. C., GIANETTO R., WATTIAUX R., APPELMANS F. Tissue fractionation studies. 6. Intracellular distribution patterns of enzymes in rat-liver tissue. Biochem J. 1955 Aug;60(4):604–617. doi: 10.1042/bj0600604. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Erlandson R. A., de Harven E. The ultrastructure of synchronized HeLa cells. J Cell Sci. 1971 Mar;8(2):353–397. doi: 10.1242/jcs.8.2.353. [DOI] [PubMed] [Google Scholar]
- FLECK A., MUNRO H. N. The precision of ultraviolet absorption measurements in the Schmidt-Thannhauser procedure for nucleic acid estimation. Biochim Biophys Acta. 1962 May 14;55:571–583. doi: 10.1016/0006-3002(62)90836-3. [DOI] [PubMed] [Google Scholar]
- GLOCK G. E., MCLEAN P. The determination of oxidized and reduced diphosphopyridine nucleotide and triphosphopyridine nucleotide in animal tissues. Biochem J. 1955 Nov;61(3):381–388. doi: 10.1042/bj0610381. [DOI] [PMC free article] [PubMed] [Google Scholar]
- HOGEBOOM G. H., SCHNEIDER W. C. Cytochemical studies. VI. The synthesis of diphosphopyridine nucleotide by liver cell nuclei. J Biol Chem. 1952 May;197(2):611–620. [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Libert J., Tondeur M., Van Hoof F. The use of conjunctival biopsy and enzyme analysis in tears for the diagnosis of homozygotes and heterozygotes with Fabry disease. Birth Defects Orig Artic Ser. 1976;12(3):221–239. [PubMed] [Google Scholar]
- Lopez-Saura P., Trouet A., Tulkens P. Analytical fractionation of cultured hepatoma cells (HTC cells). Biochim Biophys Acta. 1978 Nov 1;543(4):430–449. doi: 10.1016/0304-4165(78)90298-2. [DOI] [PubMed] [Google Scholar]
- Martin D. W., Jr, Tomkins G. M., Bresler M. A. Control of specific gene expression examined in synchronized mammalian cells. Proc Natl Acad Sci U S A. 1969 Jul;63(3):842–849. doi: 10.1073/pnas.63.3.842. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Peeters-Joris C., Vandevoorde A. M., Baudhuin P. Subcellular localization of superoxide dismutase in rat liver. Biochem J. 1975 Jul;150(1):31–39. doi: 10.1042/bj1500031. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Quintart J., Baudhuin P. Isopycnic equilibration of hepatoma-cell homogenates at different times during the cell cycle [proceedings]. Biochem Soc Trans. 1977;5(4):1174–1176. doi: 10.1042/bst0051174. [DOI] [PubMed] [Google Scholar]
- Quintart J., Baudhuin P. Uptake of horseradish peroxidase during the cell cycle of hepatoma cells in culture. Arch Int Physiol Biochim. 1976 Apr;84(2):409–410. [PubMed] [Google Scholar]
- Quintart J., Houyet M. A., Baudhuin P. Biochemical and morphological characterization of hepatoma cells during the cell cycle. Arch Int Physiol Biochim. 1975 Dec;83(5):989–990. [PubMed] [Google Scholar]
- ROBBINS E., GONATAS N. K. HISTOCHEMICAL AND ULTRASTRUCTURAL STUDIES ON HELA CELL CULTURES EXPOSED TO SPINDLE INHIBITORS WITH SPECIAL REFERENCE TO THE INTERPHASE CELL. J Histochem Cytochem. 1964 Sep;12:704–711. doi: 10.1177/12.9.704. [DOI] [PubMed] [Google Scholar]
- Salb J. M., Marcus P. I. Translational inhibition in mitotic HeLa cells. Proc Natl Acad Sci U S A. 1965 Nov;54(5):1353–1358. doi: 10.1073/pnas.54.5.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Samuels H. H., Tomkins G. M. Relation of steroid structure to enzyme induction in hepatoma tissue culture cells. J Mol Biol. 1970 Aug 28;52(1):57–74. doi: 10.1016/0022-2836(70)90177-4. [DOI] [PubMed] [Google Scholar]
- Sauvage P., Trouet A., Tulkens P. Purification and antigenic characterization of plasma membrane from hepatoma cells in culture (HTC cells). Arch Int Physiol Biochim. 1975 Dec;83(5):995–996. [PubMed] [Google Scholar]
- Sellers L., Granner D. Regulation of tyrosine aminotrasferase activity in two liver-derived permanent cell lines. J Cell Biol. 1974 Feb;60(2):337–345. doi: 10.1083/jcb.60.2.337. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Trouet A. Perspectives in cancer research. Increased selectivity of drugs by linking to carriers. Eur J Cancer. 1978 Feb;14(2):105–111. doi: 10.1016/0014-2964(78)90167-6. [DOI] [PubMed] [Google Scholar]
- Tulkens P., Beaufay H., Trouet A. Analytical fractionation of homogenates from cultured rat embryo fibroblasts. J Cell Biol. 1974 Nov;63(2 Pt 1):383–401. doi: 10.1083/jcb.63.2.383. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WURTMAN R. J., AXELROD J. A SENSITIVE AND SPECIFIC ASSAY FOR THE ESTIMATION OF MONOAMINE OXIDASE. Biochem Pharmacol. 1963 Dec;12:1439–1441. doi: 10.1016/0006-2952(63)90215-6. [DOI] [PubMed] [Google Scholar]
- Warburton M. J., Wynn C. H. Characterisation of the lysosomal heterogeneity in Chinese-hamster fibroblasts. Eur J Biochem. 1976 Jun 1;65(2):341–348. doi: 10.1111/j.1432-1033.1976.tb10347.x. [DOI] [PubMed] [Google Scholar]
- Warmsley A. M., Phillips B., Pasternak C. A. The use of zonal centrifugation to study membrane formation during the life cycle of mammalian cells. Synthesis of 'marker' enzymes and other components of cellular organelles. Biochem J. 1970 Dec;120(4):683–688. doi: 10.1042/bj1200683. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wibo M., Amar-Costesec A., Berthet J., Beaufay H. Electron microscope examination of subcellular fractions. 3. Quantitative analysis of the microsomal fraction isolated from rat liver. J Cell Biol. 1971 Oct;51(1):52–71. doi: 10.1083/jcb.51.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Duve C., de Barsy T., Poole B., Trouet A., Tulkens P., Van Hoof F. Commentary. Lysosomotropic agents. Biochem Pharmacol. 1974 Sep 15;23(18):2495–2531. doi: 10.1016/0006-2952(74)90174-9. [DOI] [PubMed] [Google Scholar]