Conserved ETS domain arginines mediate DNA binding, nuclear localization, and a novel mode of bZIP interaction.
The DNA-binding ETS transcription factor Spi-1/PU.1 is of central importance in determining the myeloid-erythroid developmental switch and is required for monocyte and osteoclast differentiation. Many monocyte genes are dependent upon this factor, including the gene that codes for interleukin-1beta. It has long been known that the conserved ETS DNA-binding domain of Spi-1/PU.1 functionally cooperates via direct association with a diverse collection of DNA-binding proteins, including members of the basic leucine zipper domain (bZIP) family. However, the molecular basis for this interaction has long been elusive. Using a combination of approaches, we have mapped a single residue on the surface of the ETS domain critical for protein tethering by the C/EBPbeta carboxyl-terminal bZIP domain. This residue is also important for nuclear localization and DNA binding. In addition, dependence upon the leucine zipper suggests a novel mode for both protein-DNA interaction and functional cooperativity.
Listman
James A
JA
SUNY Upstate Medical University, Syracuse, New York 13210, USA.
Wara-aswapati
Nawarat
N
Race
JoAnne E
JE
Blystone
Lisa W
LW
Walker-Kopp
Nancy
N
Yang
Zhiyong
Z
Auron
Philip E
PE
D001120:Arginine / Q000737:chemistry; D050976:Basic-Leucine Zipper Transcription Factors / Q000737:chemistry*; D015153:Blotting, Western; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000737:chemistry; D002412:Cations; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism*; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007375:Interleukin-1 / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism; D008954:Models, Biological; D008957:Models, Genetic; D008958:Models, Molecular; D009000:Monocytes / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry; D011518:Proto-Oncogene Proteins / Q000737:chemistry; D015534:Trans-Activators / Q000737:chemistry; D015533:Transcriptional Activation; D000099232:Proto-Oncogene Protein Spi-1
D001120:Arginine; D050976:Basic-Leucine Zipper Transcription Factors; D022782:CCAAT-Enhancer-Binding Protein-beta; D002412:Cations; D004247:DNA; D005982:Glutathione Transferase; D007375:Interleukin-1; D008156:Luciferases; D011506:Proteins; D011518:Proto-Oncogene Proteins; D015534:Trans-Activators; D000099232:Proto-Oncogene Protein Spi-1