Monoclonal antibodies to NF-Y define its function in MHC class II and albumin gene transcription.
NF-Y is a sequence-specific DNA-binding protein which, as a heterodimer, recognizes CCAAT motifs in a variety of transcriptional promoters. We have generated a panel of monoclonal and affinity-purified polyclonal antibodies directed against various epitopes of NF-Y. These reagents are highly specific for either of the A or B subunits; we have mapped the epitopes recognized by the monoclonal antibodies to the glutamine-rich activation domain of NF-YA. The antibodies inhibit in vitro transcription from the promoters of the albumin gene and of Ea, a class II gene of the major histocompatibility complex. These data definitively demonstrate the role of NF-Y in regulating the transcription of two tissue-specific genes whose expression patterns do not overlap. Interestingly, the antibodies cannot inhibit a formed pre-initiation complex, but do block reinitiation of subsequent rounds of transcription from the same templates.
Mantovani
R
R
Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, France.
Pessara
U
U
Tronche
F
F
Li
X Y
XY
Knapp
A M
AM
Pasquali
J L
JL
Benoist
C
C
Mathis
D
D
D000418:Albumins / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000915:Antibody Affinity; D000918:Antibody Specificity; D001483:Base Sequence; D022762:CCAAT-Enhancer-Binding Proteins; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000276:immunology / Q000502:physiology*; D000939:Epitopes; D005786:Gene Expression Regulation; D005802:Genes, MHC Class II; D051379:Mice; D008807:Mice, Inbred BALB C; D008957:Models, Genetic; D008969:Molecular Sequence Data; D014157:Transcription Factors / Q000276:immunology / Q000502:physiology*; D014158:Transcription, Genetic
D000418:Albumins; D000911:Antibodies, Monoclonal; D022762:CCAAT-Enhancer-Binding Proteins; D004268:DNA-Binding Proteins; D000939:Epitopes; D014157:Transcription Factors