scholarly journals Studies on the high-mobility-group non-histone proteins from hen oviduct

1979 ◽  
Vol 181 (3) ◽  
pp. 585-591 ◽  
Author(s):  
C S Teng ◽  
G K Andrews ◽  
C T Teng
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
High Mobility ◽  
High Mobility Group ◽  
Micrococcal Nuclease ◽  
Hmg Proteins ◽  
Deoxyribonuclease I ◽  
Micrococcal Nuclease Digestion ◽  
Calf Thymus ◽  
Nuclease Digestion ◽  
Limited Digestion

Nuclear high-mobility-group (HMG) proteins were isolated from hen oviduct. These were proteins HMG-1, −2, −3, −14 and −17, which are equivalent to the classification of calf thymus HMG proteins. Hen oviduct proteins HMG-1 and −2 were individually isolated by HCIO4.extraction and CM-Sephadex chromatographic separation. Their mol.wts. were determined as 28 000 and 27 000, respectively. The proteins have a high content of acidic and basic amino acids. The association of proteins HMG-1 and −2 with the genome of hen oviduct nuclei was probed by a limited digestion with nucleases. Hen oviduct nuclei were incubated with deoxyribonuclease I or micrococcal nuclease until 10% of the DNA was digested. The nuclear suspension was centrifuged and the contents of proteins HMG-1 and −2 in the supernatant and sediment fractions were analysed by polyacrylamide-gel electrophoresis. HMG proteins were found to be preferentially released by micrococcal-nuclease digestion rather than by deoxyribonuclease I.

A study of the localization of high mobility group proteins in chromatin

1978 ◽  
Vol 56 (6) ◽  
pp. 480-491 ◽  
Author(s):  
Beatriz Levy W. ◽  
Gordon H. Dixon
Keyword(s):  
Mouse Brain ◽  
High Mobility ◽  
Cell Types ◽  
Dnase I ◽  
High Mobility Group ◽  
Micrococcal Nuclease ◽  
Hmg Proteins ◽  
Calf Thymus ◽  
Limited Digestion ◽  
Active Genes

High mobility group (HMG) proteins from fetal calf thymus and mouse brain chromatin were purified and compared electrophoretically. The four major HMG proteins characteristic of fetal calf thymus chromatin (HMG's 1, 2, 14, and 17) were also found to be present in mouse brain chromatin.Nuclei from these two eucaryotic tissues were digested with DNase I and micrococcal nuclease and the acid-soluble proteins solubilized by the two nucleases in both tissues were analyzed on starch gels.Limited digestion of fetal calf thymus nuclei with DNase I led to the solubilization of a substantial fraction of proteins HMG-1 and HMG-2 together with smaller amounts of H1. In addition, limited digestion with micrococcal nuclease released approximately 70% of HMG's 1 and 2 and variable amounts of H1 into the soluble fraction. The observation that HMG proteins 1 and 2 are selectively solubilized under conditions in which active genes have been shown to be preferentially digested in various other cell types suggests their selective association with chromatin regions which are transcriptionally competent.

scholarly journals Calcium-dependent ADP-ribosylation of high-mobility-group I (HMGI) proteins

1996 ◽  
Vol 317 (3) ◽  
pp. 865-870 ◽  
Author(s):  
Vincenzo GIANCOTTI ◽  
Antonella BANDIERA ◽  
Chiara SINDICI ◽  
Laura PERISSIN ◽  
Colyn CRANE-ROBINSON
Keyword(s):  
High Mobility ◽  
High Mobility Group ◽  
Micrococcal Nuclease ◽  
Calcium Dependent ◽  
Lung Carcinoma Cells ◽  
Group I ◽  
Micrococcal Nuclease Digestion ◽  
Nuclease Digestion ◽  
Full Complement ◽  
Lewis Lung Carcinoma Cells

Micrococcal nuclease digestion of nuclei from mouse Lewis lung carcinoma cells releases a protein mixture into the supernatant that lacks histone H1 and contains a full complement of high-mobility-group I (HMGI) proteins (i.e. I, Y and I-C). This implies that all three HMGI proteins are localized at the nuclease-sensitive regions of active chromatin. It is also shown that if Ca2+ ions are present in the nuclear incubation buffer (with or without exogenous nuclease), all three HMGI proteins become ADP-ribosylated. We propose that this modification of HMGI family proteins is part of the general poly(ADP-ribosyl)ation that accompanies DNA damage in apoptosis and other processes.

scholarly journals High mobility group proteins of amphibian oocytes: a large storage pool of a soluble high mobility group-1-like protein and involvement in transcriptional events.

1983 ◽  
Vol 97 (3) ◽  
pp. 838-848 ◽  
Author(s):  
J A Kleinschmidt ◽  
U Scheer ◽  
M C Dabauvalle ◽  
M Bustin ◽  
W W Franke
Keyword(s):  
Rana Temporaria ◽  
Peptide Mapping ◽  
Gradient Centrifugation ◽  
High Mobility ◽  
High Mobility Group ◽  
Monomeric Form ◽  
Hmg Proteins ◽  
Amphibian Species ◽  
Large Oocyte ◽  
Calf Thymus

Oocytes of several amphibian species (Xenopus laevis, Rana temporaria, and Pleurodeles waltlii) contained a relatively large pool of nonchromatin-bound, soluble high mobility group (HMG) protein with properties similar to those of calf thymus proteins HMG-1 and HMG-2 (protein HMG-A; A, amphibian). About half of this soluble HMG-A was located in the nuclear sap, the other half was recovered in enucleated ooplasms. This protein was identified by its mobility on one- and two-dimensional gel electrophoresis, by binding of antibodies to calf thymus HMG-1 to polypeptides electrophoretically separated and blotted on nitrocellulose paper, and by tryptic peptide mapping of radioiodinated polypeptides. Most, if not all, of the HMG-A in the soluble nuclear protein fraction, preparatively defined as supernatant obtained after centrifugation at 100,000 g for 1 h, was in free monomeric form, apparently not bound to other proteins. On gel filtration it eluted with a mean peak corresponding to an apparent molecular weight of approximately 25,000; on sucrose gradient centrifugation it appeared with a very low S value (2-3 S), and on isoelectric focusing it appeared in fractions ranging from pH approximately 7 to 9. This soluble HMG-A was retained on DEAE-Sephacel but could be eluted already at moderate salt concentrations (0.2 M KCl). In oocytes of various stages of oogenesis HMG-A was accumulated in the nucleus up to concentrations of approximately 14 ng per nucleus (in Xenopus), corresponding to approximately 0.2 mg/ml, similar to those of the nucleosomal core histones. This nuclear concentration is also demonstrated using immunofluorescence microscopy. When antibodies to bovine HMG-1 were microinjected into nuclei of living oocytes of Pleurodeles the lateral loops of the lampbrush chromosomes gradually retracted and the whole chromosomes condensed. As shown using electron microscopy of spread chromatin from such injected oocyte nuclei, this process of loop retraction was accompanied by the appearance of variously-sized and irregularly-spaced gaps within transcriptional units of chromosomal loops but not of nucleoli, indicating that the transcription of non-nucleolar genes was specifically inhibited by this treatment and hence involved an HMG-1-like protein. These data show that proteins of the HMG-1 and -2 category, which are usually chromatin-bound components, can exist, at least in amphibian oocytes, in a free soluble monomeric form, apparently not bound to other molecules. The possible role of this large oocyte pool of soluble HMG-A in early embryogenesis is discussed as well as the possible existence of soluble HMG proteins in other cells.

scholarly journals HMG (high-mobility-group)-14/17-like proteins in calf thyroid. Thyrotropin-dependent phosphorylation and comparison with calf thymus proteins

1983 ◽  
Vol 215 (3) ◽  
pp. 643-649 ◽  
Author(s):  
E Cooper ◽  
S W Spaulding
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
High Mobility ◽  
High Mobility Group ◽  
Group 14 ◽  
Sephadex Column ◽  
High Mobility Group Proteins ◽  
Electrophoretic Mobilities ◽  
Calf Thymus ◽  
Thymus Tissue

Two-dimensional polyacrylamide-gel electrophoresis of acid extracts of thyroid and thymus tissue, and of thyroid nuclei, revealed the presence of three HClO4-soluble nuclear proteins, PS.1, PS.2 and PS.3, whose electrophoretic mobilities closely resembled those of HMG (high-mobility-group) proteins 14 and 17. PS.1 co-migrated with HMG 14 on CM-Sephadex column chromatography. Like HMG 14, PS.2 and PS.3 were phosphorylated in calf thyroid slices; 32P-labelling of PS.3 was stimulated by thyrotropin. Thyrotropin also induced a rapid increase in the labelling of A5, an HMG-14/17-like protein found in whole calf thyroid and thymus tissue, but not in thyroid nuclei.

scholarly journals Immuno-biochemical studies of a non-histone chromosomal protein in embryonic and mature chick oviduct

1980 ◽  
Vol 185 (1) ◽  
pp. 169-175 ◽  
Author(s):  
C T Teng ◽  
C S Teng
Keyword(s):  
Chick Embryo ◽  
Complement Fixation ◽  
High Mobility ◽  
High Mobility Group ◽  
Chromosomal Protein ◽  
High Mobility Group Proteins ◽  
Biochemical Studies ◽  
Nuclease Digestion ◽  
Antibody Induction ◽  
Limited Digestion

A non-histone chromosomal 95K protein (of mol.wt. 95 000) from hen oviduct was isolated and purified for antibody induction in the rabbit. Immuno-micro-complement-fixation and biochemical techniques were used to probe the presence of 95K protein in the oviduct chromatin of the embryonic and immature chick and the hen. The antiserum against 95K protein did not react with high-mobility-group proteins 1 and 2 obtained from oviduct, brain and liver, nor with histones. After limited digestion of chromatin with nucleases, until 10% DNA was hydrolysed, a small amount of 95K protein was released. Thus the 95K protein is probably not located in the region of chromatin that is sensitive to nuclease digestion. The amount of 95K protein in immature chick oviduct chromatin is less than that in the mature hen oviduct. However, the amount of 95K protein in the immature chick oviduct was increased after oestrogen administration. The presence of 95K protein in embryonic oviduct was detected in the 10-, 12-, 15- and 18- day chick embryo. The quantity of this protein increased with the age of the embryo and reached its highest value in the chromatin of the hen oviduct.

scholarly journals Tetrahymena contain two distinct and unusual high mobility group (HMG)-like proteins.

1987 ◽  
Vol 104 (6) ◽  
pp. 1485-1494 ◽  
Author(s):  
I G Schulman ◽  
R G Cook ◽  
R Richman ◽  
C D Allis
Keyword(s):  
Polyclonal Antibodies ◽  
High Mobility ◽  
Histone H1 ◽  
High Mobility Group ◽  
Hmg Proteins ◽  
Terminal Sequence ◽  
Amino Terminal ◽  
Calf Thymus ◽  
Specific Induction ◽  
Amino Terminal Sequence

Previous studies have described the existence of high mobility group (HMG)-like proteins in macronuclei of the ciliated protozoan, Tetrahymena thermophila (Hamana, K., and K. Iwai, 1979, J. Biochem. [Tokyo], 69:1097-1111; Levy-Wilson, B., M. S. Denker, and E. Ito, 1983, Biochemistry, 22:1715-1721). In this report, two of these proteins, LG-1 and LG-2, have been further characterized. Polyclonal antibodies raised against LG-1 and LG-2 fail to cross react with each other or any other macronuclear polypeptide in immunoblotting analyses. As well, LG-1 and LG-2 antibodies do not react with calf thymus, chicken, or yeast HMG proteins. Consistent with these results, a 47 amino-terminal sequence of LG-1 has been determined that shows limited homology to both calf thymus HMGs 1 and 2 and HMGs 14 and 17. Two internal sequences of V8 protease-generated peptides from LG-2 have been determined, and these do not share any homology to the LG-1 sequence or any other sequenced HMG proteins. Comparison of the partial sequences of LG-1 and LG-2 with the complete amino acid sequence of the Tetrahymena histone H1 (Wu, M., C. D. Allis, R. Richman, R. G. Cook, and M. A. Gorovsky, 1986, Proc. Natl. Acad. Sci. USA, 83:8674-8678) rules out the possibility that LG-1 and LG-2 are proteolytically derived from H1, the other major macronuclear perchloric acid-soluble protein. Interestingly, however, both LG-1 and LG-2 are efficiently extracted from macronuclei by elutive intercalation (Schröter, H., G. Maier, H. Ponsting, and A. Nordheim, 1985, Embo (Eur. Mol. Biol. Organ.) J., 4:3867-3872), suggesting that both may share yet undetermined properties with HMGs 14 and 17 of higher eukaryotes. Examination of the pattern of LG-1 and LG-2 synthesis during the sexual phase of the life cycle, conjugation, demonstrates that the synthesis of LG-1 and LG-2 is coordinately increased from basal levels during the differentiation of new macronuclei (7-13 h), suggesting that both of these proteins play a role in determining a macronuclear phenotype. However, a specific induction of LG-2 synthesis is detected in early stages of conjugation (meiotic prophase, 1-4 h), leading to maximal synthesis of LG-2 at 3 h. Interestingly, the early induction of LG-2 synthesis closely parallels the hyperphosphorylation of histone H1. Taken together, these data suggest that LG-1 and LG-2 are not strongly related to each other or to higher eukaryotic HMG proteins.(ABSTRACT TRUNCATED AT 400 WORDS)

scholarly journals Studies on the tissue specificity of the high-mobility-group non-histone chromosomal proteins from calf

1978 ◽  
Vol 173 (2) ◽  
pp. 497-505 ◽  
Author(s):  
A Rabbani ◽  
G H Goodwin ◽  
E W Johns
Keyword(s):  
Amino Acid ◽  
Gel Electrophoresis ◽  
Trichloroacetic Acid ◽  
Tissue Specificity ◽  
Polyacrylamide Gel Electrophoresis ◽  
High Mobility ◽  
Acid Precipitation ◽  
High Mobility Group ◽  
Chromosomal Proteins ◽  
Calf Thymus

The high-mobility-group (HMG) non-histone chromosomal proteins from calf thymus, liver, spleen and kidney were extracted, and fractionated by CM-Sephadex chromatography and trichloroacetic acid precipitation. The isolated proteins HMG 1, HMG 2 and HMG 17 from the tissues were compared by polyacrylamide-gel electrophoresis, isoelectric focusing and amino acid analysis. The results show that the three proteins are very similar in the tissues studied, implying a lack of tissue specificity.

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