The class III POU factor Brn-4 interacts with other class III POU factors and the heterogeneous nuclear ribonucleoprotein U.
The class III POU proteins are expressed throughout the central nervous system, including the hypothalamus, where they are often co-localized. Presumably, these POU proteins (Brain-1, Brain-2, Brain-4 and SCIP) serve as transcriptional transactivators. That they are co-expressed in some neurons suggests that, if they were to form homomeric and heteromeric complexes with each other, depending on the particular combination, they might have different DNA-binding specificities and, thus, activate different genes. We used purified fusion proteins of the four class III POU proteins in far-western assays to show that the proteins can interact. We confirmed their interactions using a two-hybrid system. Both techniques indicate that the interaction occurs through the POU domain. The far-western technique also allowed us to identify a 120-kDa nuclear protein that interacts with Brain-4. Subsequent affinity purification and microsequencing identified the protein as the heterogeneous nuclear ribonucleoprotein U (hnRNP U). This result suggests another mechanism by which a POU protein can influence gene expression: by facilitating the processing of pre-mRNA whose transcription it has stimulated.
Malik
K F
KF
Laboratory of Cell Biology, National Institute of Mental Health, National Institutes of Health, Bethesda, MD 20892, USA.
Jaffe
H
H
Brady
J
J
Young
W S
WS
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D002467:Cell Nucleus / Q000378:metabolism; D004268:DNA-Binding Proteins; D015870:Gene Expression; D005982:Glutathione Transferase; D034722:Heterogeneous-Nuclear Ribonucleoprotein U; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D007031:Hypothalamus / Q000378:metabolism; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins; D009420:Nervous System / Q000378:metabolism*; D009842:Oligopeptides; D050809:POU Domain Factors; D010446:Peptide Fragments / Q000737:chemistry; D010455:Peptides; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D055503:Protein Multimerization; D012322:RNA Precursors / Q000378:metabolism; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012261:Ribonucleoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques
D004268:DNA-Binding Proteins; D034722:Heterogeneous-Nuclear Ribonucleoprotein U; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D009419:Nerve Tissue Proteins; D009842:Oligopeptides; D050809:POU Domain Factors; D010446:Peptide Fragments; D010455:Peptides; C484088:Pou3f4 protein, rat; D012322:RNA Precursors; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; D014157:Transcription Factors; C085706:FLAG peptide; D005982:Glutathione Transferase