NUCLEIC ACIDS, PROTEIN SYNTHESIS, AND MOLECULAR GENETICS
Biochemical Characterization of the NF-Y Transcription Factor Complex during B Lymphocyte Development*

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The transcription factor, NF-Y, plays a critical role in tissue-specific major histocompatibility complex class II gene transcription. In this report the biochemical properties of the heterotrimeric NF-Y complex have been characterized during stage-specific B-cell development, and in several class II mutant B-cell lines, which represent distinct bare lymphocyte syndrome class II genetic complementation groups. The NF-Y complex derived from class II+ mature B-cells bound with high affinity to anion exchangers, and eluted as an intact trimeric complex, whereas, NF-Y derived from class IIplasma B-cells, and from bare lymphocyte syndrome group II cell lines, RJ2.2.5 and RM3, dissociated into discrete NF-YA and NF-YB:C subunit fractions. Recombination of the MPC11 plasma B-cell derived NF-Y A:B:C complex with the low molecular mass protein fraction, NF-Y-associated factors (YAFs), derived from mature A20 B-cell nuclei, conferred high affinity anion exchange binding to NF-Y as an intact trimeric complex. Recombination of the native NF-YA:B:C complex with the transcriptional cofactor, PC4, likewise conferred high affinity NF-Y binding to anion exchangers, and stabilized NF-Y interaction with CCAAT-box DNA motifs in vitro. Interaction between PC4 and NF-Y was mapped to the C-terminal region of PC4, and the subunit interaction subdomain of the highly conserved DNA binding-subunit interaction domain (DBD) of NF-YA. These results suggest that in class II+ mature B-cells NF-Y is associated with the protein cofactor, PC4, which may play an important role in NF-Y-mediated transcriptional control of class II genes.

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This work was supported by United States Public Health Service Grant AI37686 from NIAID, National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.