Protein acetylation regulates both PU.1 transactivation and Ig kappa 3' enhancer activity.
Igkappa gene expression and chromatin structure change during B cell development. At the pre-B cell stage, the locus is relatively hypoacetylated on histone H3, whereas it is hyperacetylated at the plasma cell stage. We find in this study that the histone deacetylase inhibitor, trichostatin A (TSA) stimulated 3' enhancer activity through the PU.1 binding site. TSA also stimulated PU.1 transactivation potential. PU.1 activity was increased by the coactivator acetyltransferase protein, p300, and p300 physically interacted with PU.1 residues 7-30. PU.1 served as a substrate for p300 and was acetylated on lysine residues 170, 171, 206, and 208. Mutation of PU.1 lysines 170 and 171 did not affect PU.1 DNA binding, but did lower the ability of PU.1 to activate transcription in association with p300. Lysine 170 was acetylated in pre-B cells and plasmacytoma cells, but TSA treatment did not stimulate PU.1 acetylation at this residue arguing that a second mechanism can stimulate 3' enhancer activity. Using chromatin immunoprecipitation assays we found that TSA caused preferential acetylation of histone H3 at the 3' enhancer. The relevance of these studies for PU.1 function in transcription and hemopoietic development is discussed.
Bai
Yuchen
Y
Department of Animal Biology, University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA 19104, USA.
Srinivasan
Lakshmi
L
Perkins
Leslie
L
Atchison
Michael L
ML
D020413:3' Untranslated Regions / Q000378:metabolism*; D000107:Acetylation; D000595:Amino Acid Sequence; D000818:Animals; D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D004742:Enhancer Elements, Genetic / Q000502:physiology*; D051548:Histone Acetyltransferases / Q000378:metabolism / Q000502:physiology; D006657:Histones / Q000378:metabolism*; D006877:Hydroxamic Acids / Q000494:pharmacology; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D014157:Transcription Factors / Q000378:metabolism; D015533:Transcriptional Activation; D050880:p300-CBP Transcription Factors; D000099232:Proto-Oncogene Protein Spi-1; D000099224:p300-CBP-Associated Factor
D020413:3' Untranslated Regions; D018797:Cell Cycle Proteins; D051548:Histone Acetyltransferases; D006657:Histones; D006877:Hydroxamic Acids; D007145:Immunoglobulin kappa-Chains; D011518:Proto-Oncogene Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D050880:p300-CBP Transcription Factors; D000099232:Proto-Oncogene Protein Spi-1; D000099224:p300-CBP-Associated Factor; C012589:trichostatin A