Chromosomal protein poly(ADP-ribosyl)ation in pancreatic nucleosomes

1982 ◽  
Vol 60 (3) ◽  
pp. 295-305 ◽  
Author(s):  
R. J. Aubin ◽  
V. T. Dam ◽  
J. Miclette ◽  
Y. Brousseau ◽  
G. G. Poirier
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
High Mobility ◽  
Histone H1 ◽  
Polymerase Activity ◽  
Chromosomal Protein ◽  
Chromosomal Proteins ◽  
Adp Ribosylation ◽  
The Core ◽  
Lithium Dodecyl Sulfate ◽  
Dodecyl Sulfate

When pancreatic chromatin fragments were prepared and resolved in the presence of 80 mM NaCl, endogenous poly(ADP-ribose) polymerase activity was found to be maximal in nucleosome periodicities of four to five units and did not respond to any further increases in nucleosomal architecture. Furthermore, in nucleosome complexities spanning 1 through 14 and over unit lengths, polyacrylamide gel electrophoresis on acid–urea and acid–urea–Triton gels has shown pancreatic histone H1 to be the only actively ADP-ribosylated histone species. The extent of ADP-ribosylation of histone H1 was also demonstrated to retard the protein's mobility in acid–urea, acid–urea–Triton, and lithium dodecyl sulfate polyacrylamide gels and to consist of at least 12 distinct ADP-ribosylated species extractable in all nucleosome complexities studied. Finally, extraction and subsequent electrophoresis of total chromosomal proteins in the presence of lithium dodecyl sulfate also evidenced heavy ADP-ribosylation at the level of nonhistone chromosomal proteins of the high mobility group comigrating in the core histone region, as well as in the topmost region of the gels where poly(ADP-ribose) polymerase was found to form a poly(ADP-ribosyl)ated aggregate.

ADP-ribosylation of histones and acid-soluble, high mobility group like chromosomal proteins from Physarum polycephalum

1987 ◽  
Vol 65 (1) ◽  
pp. 81-85 ◽  
Author(s):  
G. G. Poirier ◽  
Serge Coté ◽  
Dominic Pallotta
Keyword(s):  
Gel Electrophoresis ◽  
Physarum Polycephalum ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein A ◽  
High Mobility ◽  
High Mobility Group ◽  
Chromosomal Proteins ◽  
Cellular Slime Mould ◽  
Adp Ribosylation ◽  
Cellular Slime

Nuclei from the cellular slime mould Physarum polycephalum were incubated with [32P]NAD. The chromosomal basic proteins were acid extracted and separated by two-dimensional polyacrylamide gel electrophoresis. After autoradiography the poly(ADP-ribosylated) proteins were identified. Histone H1 was the major acceptor. Histones H2B and H2A were significantly modified, although to a lesser extent than H1. In addition, the acid-soluble, high mobility group-like proteins AS-2 and AS-3 and the protein A-24 showed some modification. Histones H3 and H4 were not modified. The pattern of ADP-ribosylation did not change with NAD concentrations between 1 and 100 μM NAD.

scholarly journals Does high-mobility-group non-histone protein HMG 1 interact specifically with histone H1 subfractions?

1979 ◽  
Vol 183 (3) ◽  
pp. 657-662 ◽  
Author(s):  
P D Cary ◽  
K V Shooter ◽  
G H Goodwin ◽  
E W Johns ◽  
J Y Olayemi ◽  
...  
Keyword(s):  
Specific Interaction ◽  
High Mobility ◽  
Histone H1 ◽  
High Mobility Group ◽  
Chromosomal Protein ◽  
Histone Protein ◽  
Total Histone ◽  
Equilibrium Sedimentation ◽  
Histone H5

The interaction of the non-histone chromosomal protein HMG (high-mobility group) 1 with histone H1 subfractions was investigated by equilibrium sedimentation and n.m.r. sectroscopy. In contrast with a previous report [Smerdon & Isenberg (1976) Biochemistry 15, 4242–4247], it was found, by using equilibrium-sedimentation analysis, that protein HMG 1 binds to all three histone H1 subfractions CTL1, CTL2, and CTL3, arguing against there being a specific interaction between protein HMG 1 and only two of the subfractions, CTL1 and CTL2. Raising the ionic strength of the solutions prevents binding of protein HMG 1 to total histone H1 and the three subfractions, suggesting that the binding in vitro is simply a non-specific ionic interaction between acidic regions of the non-histone protein and the basic regions of the histone. Protein HMG 1 binds to histone H5 also, supporting this view. The above conclusions are supported by n.m.r. studies of protein HMG 1/histone H1 subfraction mixtures. When the two proteins were mixed, there was little perturbation of the n.m.r. spectra and there was no evidence for specific interaction of protein HMG 1 with any of the subfractions. It therefore remains an open question as to whether protein HMG 1 and histone H1 are complexed together in chromatin.

Isolation and characterization of histones and other acid-soluble chromosomal proteins from Physarum polycephalum

1982 ◽  
Vol 60 (3) ◽  
pp. 263-271 ◽  
Author(s):  
Serge Côté ◽  
Paul Nadeau ◽  
James M. Neelin ◽  
Dominick Pallotta
Keyword(s):  
Amino Acid ◽  
Physarum Polycephalum ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
High Mobility ◽  
Apparent Molecular Weight ◽  
Chromosomal Protein ◽  
Basic Proteins ◽  
Isolation And Characterization ◽  
Core Histones

Chromosomal basic proteins were isolated from amoebal and plasmodial stages of the acellular slime mold Physarum polycephalum. Polyacrylamide electrophoresis on high resolution acid–urea gels separated the five histone fractions in the sequence H1, H2A, H2B, H3, andH4. Under these electrophoretic conditions Physarum histones migrated more like plant (rye) than animal (calf) histones. Furthermore, Physarum histones H1, H2A, and H2B have higher molecular weights on sodium dodecyl sulfate (SDS) gels than the corresponding calf fractions. No differences were detected between amoebal and plasmodial histones on either acid–urea or SDS–polyacrylamide gel electrophoresis. Amoebal basic proteins were fractionated by exclusion chromatography. The five histone fractions plus another major acid-soluble chromosomal protein (AS) were isolated. The Physarum core histones had amino acid compositions more closely resembling those of the calf core histones than of rye, yeast, or Dictyostelium. Although generally similar in composition to the plant and animal H1 histones, the Physarum H1 had a lower lysine content. The AS protein was extracted with 5% perchloric acid or 0.5 M NaCl, migrated between histones H3 and H4 on acid–urea polyacrylamide gels, and had an apparent molecular weight of 15 900 on SDS gels. It may be related to a protein migrating near H1. Both somewhat resembled the high mobility group proteins in amino acid composition.

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.

ADP-ribosylation of pancreatic histone H1 and of other histones

1980 ◽  
Vol 58 (6) ◽  
pp. 509-515 ◽  
Author(s):  
G. G. Poirier ◽  
P. Savard
Keyword(s):  
Column Chromatography ◽  
Salt Concentration ◽  
Gel Electrophoresis ◽  
Histone H1 ◽  
Adp Ribosylation ◽  
The Core ◽  
Core Histones ◽  
Differential Binding

Incubation of pancreatic nuclei with high NAD concentrations resulted in increased ADP-ribosylation of histone H1. Interaction of [3H]ADP-ribosylated histone H1 with chromatin was significantly different from unmodified histone H1. The presence of a protein which is eluted at a lower salt concentration and which is ADP-ribosylated was also noticed. Pancreatic histones were isolated by column chromatography and their degree of ADP-ribosylation evaluated both by gel electrophoresis and by chromatography: histone H1 was the main acceptor while the core histones H3, H2B, and H2A were lightly labelled. Histones H1 and H10 have a differential binding to pancreatic chromatin and histone H10 is not ADP-ribosylated.

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