CRISP-3, a protein with homology to plant defense proteins, is expressed in mouse B cells under the control of Oct2.
The Oct2 transcription factor is expressed throughout the B-lymphoid lineage and plays an essential role during the terminal phase of B-cell differentiation. Several genes specifically expressed in B lymphocytes have been identified that contain a functional octamer motif in their regulatory elements. However, expression of only a single gene, the murine CD36 gene, has been shown to date to be dependent on Oct2. Here, we present the identification and characterization of a further gene, coding for cysteine-rich secreted protein 3 (CRISP-3), whose expression in B cells is regulated by Oct2. We show that CRISP-3 is expressed in the B-lymphoid lineage specifically at the pre-B-cell stage. By using different experimental strategies, including nuclear run-on experiments, we demonstrate that this gene is transcriptionally activated by Oct2. Furthermore, analysis of CRISP-3 expression in primary B cells derived from either wild-type or Oct2-deficient mice demonstrates the dependence on Oct2. Two variant octamer motifs were identified in the upstream promoter region of the crisp-3 gene, and Oct2 interacts with both of them in vitro. Cotransfection experiments with expression vectors for Oct1 and Oct2 together with a reporter driven by the crisp-3 promoter showed that transcriptional activation of this promoter can only be achieved with Oct2. The C-terminal transactivation domain of Oct2 is required for this activation. Finally, introducing specific mutations in the two variant octamer motifs revealed that both of them are important for full transcriptional activation by Oct2.
Pfisterer
P
P
Zentrum für Molekulare Biologie, Universität Heidelberg, Germany.
König
H
H
Hess
J
J
Lipowsky
G
G
Haendler
B
B
Schleuning
W D
WD
Wirth
T
T
D016475:3T3 Cells; D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism*; D001798:Blood Proteins / Q000737:chemistry; D002460:Cell Line; D003545:Cysteine; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000378:metabolism; D023082:Defensins; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D008221:Lymphoid Tissue / Q000378:metabolism; D008297:Male; D051379:Mice; D050813:Octamer Transcription Factor-2; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis; D012471:Salivary Proteins and Peptides / Q000096:biosynthesis* / Q000737:chemistry; D029589:Seminal Plasma Proteins; D017386:Sequence Homology, Amino Acid; D012727:Sex Characteristics; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics; D013601:T-Lymphocytes / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects*; D014162:Transfection; D014407:Tumor Cells, Cultured
D001798:Blood Proteins; C489926:CRISP3 protein, human; C489927:Crisp3 protein, mouse; D017931:DNA Primers; D004268:DNA-Binding Proteins; D023082:Defensins; D050813:Octamer Transcription Factor-2; C489206:Pou2f2 protein, mouse; D011994:Recombinant Proteins; D012471:Salivary Proteins and Peptides; D029589:Seminal Plasma Proteins; D014157:Transcription Factors; D004958:Estradiol; D003545:Cysteine