Relationship between the structure of chromosomal protein HMG1 and its accumulation in the cell nucleus.
When microinjected into the cytoplasm of cultured mammalian cells, non-histone chromosomal protein HMG1 migrates into the nucleus and binds to the chromatin. To define the features of the HMG1 molecule which are essential for this activity, fragments of HMG1 and chemically modified HMG1 molecules were injected into HeLa cells and the capacity of each of these probes to accumulate in the nucleus was measured by an autoradiographic technique. Fragments representing the C-terminal and central portions of HMG1 did not concentrate in the nucleus; a fragment which consisted of the N-terminal two-thirds of the molecule and which lacked the 41 consecutive aspartate and glutamate residues located near the C-terminal end of the molecule accumulated to about the same extent as intact HMG1. When the amino groups of HMG1 were chemically modified, there was a progressive loss in the ability of the protein to accumulate in the nucleus; derivatization of one-fourth of the total amino groups reduced the concentration of microinjected protein in the nucleus relative to that in the cytoplasm to one-half of the original value. In contrast, modification of one-fourth of the total carboxyl groups did not significantly affect the capacity of HMG1 to accumulate in the nucleus, although further modification resulted in decreased nuclear accumulation. Iodination of tyrosine residues was without effect and modification of the cysteine residues had only a modest effect on the ability of HMG1 to concentrate in the nucleus.
Kuehl
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L
Rechsteiner
M
M
Wu
L
L
D000596:Amino Acids / Q000032:analysis; D002459:Cell Fusion; D002467:Cell Nucleus / Q000378:metabolism*; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000378:metabolism; D006367:HeLa Cells / Q000378:metabolism; D006609:High Mobility Group Proteins / Q000378:metabolism*; D006801:Humans; D007457:Iodine Radioisotopes; D008970:Molecular Weight; D010222:Parainfluenza Virus 1, Human; D010446:Peptide Fragments / Q000032:analysis; D013950:Thymus Gland / Q000378:metabolism; D014357:Trypsin
D000596:Amino Acids; D006609:High Mobility Group Proteins; D007457:Iodine Radioisotopes; D010446:Peptide Fragments; D014357:Trypsin; D003488:Cyanogen Bromide