Abstract
NF-Y is a highly conserved transcription factor which recognizes CCAAT motifs in a variety of genes. We report here the genomic organization of the genes encoding both subunits, which gives interesting clues about the functional organization of the proteins. We also report the existence of isoforms of NF-YA which result from differential splicing. These alternative splicing events map within a glutamine-rich activation domain and show a marked cell- and tissue-specific bias.
Highlights
NF-Y is a highly conserved transcription factor which recognizes CCAAT motifs in a variety of genes
We report the existence of isoforms of NF-YA which result from differential splicing
NF-Y is a eukaryotic transcription factor that binds with high specificity to CCAAT motifs in the promoter regions of genes transcribed by RNA polymerase II(B)
Summary
PCRAmplijication and Library Screening-Poly(A+) RNAwas prepared from a variety of cell lines or mouse tissues, essentially as described (Koch et al, 1989). The RNA was reverse-transcribed and amplified by PCR under standard conditions, using primers mapping just outside the NF-YA coding sequences (5”CTAGGAATTCAGATCTGTAGAGTGAAGCTTCAGG-3’) and 5”CTAGGAATTCAGATCTCCCCACTGAAGTCAGTCC-3’) and incorporating artificial cloning sites. CDNA and genomic h libraries were screened according to standard protocols, using cDNA fragments from clones YA-EM13 and YB-EM38 (Hooft van Huijsduijnen et al, 1990). SI Nuclease Mapping-Relative amounts of short and long form NF-YA mRNA were determined by quantitative S1 nuclease analysis, performed as described (Koch et al, 1989). The probe was a singlestranded complementary DNA fragment from long form NF-YA, 5’ end-labeled (at position 235) and extending across the variable element and into theplasmid vector sequences. Retardation Assays-Standard or pore-gradient gel retardation assays were performed as previously described, with a 22-mer dsoligonucleotide encompassing the Y box of the Ea promoter (Hooft van Huijsduijnen et al, 1987; Dorn et al, 1987).
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
