Abstract

: The mRNA 3'-end processing reaction has been shown to play an important role in modulating Ig heavy chain poly(A) site use in B-cell development. This proposal focuses on defining the mechanisms of action of hnRNPF and H/H', auxiliary polyadenylation factors, and how they operate to influence poly(A) site choice during the induction of myeloma/plasma and lymphoma/memory B cells. We propose the following:
Aim 1. To define the role of hnRNP F in binding RNA and blocking 3' end processing in cells. A. We will test the hypothesis that specific domains of hnRNP F are responsible for its RNA binding and inhibition of 3' end processing. B. We will test the hypothesis that the differences between hnRNP F and the closely related proteins hnRNP H/H', in the carboxyl-terminal region and between RNA binding domains 2 and 3, are responsible for the differing biological effects of these molecules on 3' processing. C. We will test the hypothesis that specific phosphorylation of CTD will enhance the competition between hnRNP F and CstF-64.
AIM 2. We will test the hypothesis that differential loading of 3'-end processing factors occurs both along the Ig heavy chain gene and during different stages of B-cell development.
AIM 3. We will test the hypothesis that over-expression of hnRNP F in authentic plasma cells will suppressIg heavy chain secretion by reducing the amount of secretory specific mRNA.
AIM 4. We hypothesize that the increased expression of PC4 we have seen in plasma cells influences 3'-endformation by interacting directly with CstF-64.The experiments described in this proposal attempt to find differences in cells that may serve as markers to identify memory, naive, and plasma B-cells as well as to understand the molecular events that determine how memory an/or B-cells may be activated to differentiate into plasma cells. Understanding plasma cell development is important for vaccine development to eradicate infectious diseases, mediating allergic responses and autoimmune diseases, and limiting tumor growth. Understanding the control of polyadenylation is important for understanding and possibly controlling aspects of cell growth, differentiation and malignancy.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA086433-07
Application #
7008519
Study Section
Immunobiology Study Section (IMB)
Program Officer
Howcroft, Thomas K
Project Start
2000-02-15
Project End
2008-01-31
Budget Start
2006-02-01
Budget End
2007-01-31
Support Year
7
Fiscal Year
2006
Total Cost
$253,768
Indirect Cost

  Institution

Name
University of Pittsburgh
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213

  Publications

Park, Kyung Soo; Bayles, Ian; Szlachta-McGinn, Alec et al. (2014) Transcription elongation factor ELL2 drives Ig secretory-specific mRNA production and the unfolded protein response. J Immunol 193:4663-74
Kardava, Lela; Yang, Qi; St Leger, Anthony et al. (2011) The B lineage transcription factor E2A regulates apoptosis in chronic lymphocytic leukemia (CLL) cells. Int Immunol 23:375-84
Martincic, Kathleen; Alkan, Serkan A; Cheatle, Alys et al. (2009) Transcription elongation factor ELL2 directs immunoglobulin secretion in plasma cells by stimulating altered RNA processing. Nat Immunol 10:1102-9
Pal, Rekha; Janz, Martin; Galson, Deborah L et al. (2009) C/EBPbeta regulates transcription factors critical for proliferation and survival of multiple myeloma cells. Blood 114:3890-8
Price, Paul W; McKinney, Elizabeth C; Wang, Youliang et al. (2009) Engineered cell surface expression of membrane immunoglobulin as a means to identify monoclonal antibody-secreting hybridomas. J Immunol Methods 343:28-41
Yang, Qi; Kardava, Lela; St Leger, Anthony et al. (2008) E47 controls the developmental integrity and cell cycle quiescence of multipotential hematopoietic progenitors. J Immunol 181:5885-94
Shell, Scott A; Martincic, Kathleen; Tran, Joseph et al. (2007) Increased phosphorylation of the carboxyl-terminal domain of RNA polymerase II and loading of polyadenylation and cotranscriptional factors contribute to regulation of the ig heavy chain mRNA in plasma cells. J Immunol 179:7663-73
Borghesi, Lisa; Milcarek, Christine (2007) Innate versus adaptive immunity: a paradigm past its prime? Cancer Res 67:3989-93
Alkan, Serkan A; Martincic, Kathleen; Milcarek, Christine (2006) The hnRNPs F and H2 bind to similar sequences to influence gene expression. Biochem J 393:361-71
Shell, Scott A; Hesse, Candice; Morris Jr, Sidney M et al. (2005) Elevated levels of the 64-kDa cleavage stimulatory factor (CstF-64) in lipopolysaccharide-stimulated macrophages influence gene expression and induce alternative poly(A) site selection. J Biol Chem 280:39950-61

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