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J. Biol. Chem., Vol. 280, Issue 13, 13171-13178, April 1, 2005

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Regulation of -Fibrinogen Chain Expression by Heterogeneous Nuclear Ribonucleoprotein A1^*

Hui Xia

From the Lindsley F. Kimball Research Institute, New York Blood Center, New York, New York 10021

Earlier studies showed that HepG2 cells stably transfected with any one fibrinogen chain cDNA enhanced the expression of the other two fibrinogen chains. In this report, a regulatory element "TGCTCTC" in the -fibrinogen promoter region, -322 to -316, is identified, which is involved in increased expression of chain in HepG2 cells that are transfected with B fibrinogen cDNA. By electrophoretic mobility shift assay, three DNA-protein complexes were found to form with the regulatory element. The amount of the protein complexes that bind with the regulatory element was much reduced in HepG2 cells transfected with B cDNA. By DNA-affinity chromatography, mass spectrometry, and supershift assay, human heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) was identified as a component of the complexes. Overexpression of hnRNP A1 suppressed basal -fibrinogen transcription. These results indicate that the basal expression of -fibrinogen is regulated by a constitutive transcriptional repressor protein, hnRNP A1, and the decreased binding activity of hnRNP A1 leads to the overexpression of chain in HepG2 cells that overexpress the B chain.

Received for publication, December 15, 2004 , and in revised form, January 20, 2005.

^* This work was supported by the F. M. Kirby Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: New York Blood Center, 310 E. 67th St., New York, NY 10021. Tel.: 212-570-3342; Fax: 212-879-0243; E-mail: hxia{at}nybloodcenter.org.

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