Abstract
To investigate the in vivo functions of PU.1 and Spi-B, two highly related Ets transcription factors, we previously generated PU. 1(+/+)Spi-B(-/-) and PU.1(+/-)Spi-B(-/-) mice and demonstrated a significant decrease in B-cell receptor (BCR) signaling in mutants. Major components of BCR signaling appear to be expressed at normal levels in these mice, implying that PU.1 and Spi-B cooperate in the transcription of additional target genes important for antigen receptor signaling. We used subtractive hybridization to identify novel in vivo PU.1/Spi-B target genes and determined that the expression of a heptahelical receptor, P2Y10, is dramatically reduced in PU.1(+/-)Spi-B(-/-) B-cells. Further analysis shows that P2Y10 expression is restricted to lymphoid cells and parallels that of Spi-B in B-lymphocytes. Lastly, the P2Y10 promoter contains a PU. 1/Spi-B binding site functionally required for efficient transcription in B-cells. Thus, P2Y10 is likely to be a direct in vivo transcriptional target for PU.1 and Spi-B and provides a unique model to explore transcriptional regulation by this Ets factor subfamily. Furthermore, P2Y10 suggests an intriguing connection between heterotrimeric G-proteins and BCR signaling.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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3T3 Cells
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Amino Acid Sequence
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Animals
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B-Lymphocytes / metabolism
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Binding Sites
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Binding, Competitive
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Blotting, Northern
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Cell Lineage
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DNA / genetics
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DNA / metabolism
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DNA, Complementary / genetics
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism
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DNA-Binding Proteins / physiology*
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Gene Expression
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Mice
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Mice, Knockout
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Molecular Sequence Data
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Promoter Regions, Genetic
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Proto-Oncogene Proteins / genetics
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Proto-Oncogene Proteins / metabolism
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Proto-Oncogene Proteins / physiology*
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RNA, Messenger / genetics
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RNA, Messenger / isolation & purification
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RNA, Messenger / metabolism
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Receptors, Antigen, B-Cell / genetics*
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Receptors, Antigen, B-Cell / metabolism
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Sequence Homology, Amino Acid
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Tissue Distribution
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Trans-Activators / genetics
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Trans-Activators / metabolism
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Trans-Activators / physiology*
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Transcription Factors / genetics
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Transcription Factors / metabolism
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Transcription Factors / physiology*
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Transcription, Genetic*
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Tumor Cells, Cultured
Substances
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DNA, Complementary
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DNA-Binding Proteins
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Proto-Oncogene Proteins
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RNA, Messenger
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Receptors, Antigen, B-Cell
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Trans-Activators
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Transcription Factors
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proto-oncogene protein Spi-1
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SPIB protein, human
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DNA