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GENE EXPRESSION IN ENTAMOEBA HISTOLYTICA

Award Number: R01AI037941
ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 02/01/1996
PERIOD OF PERFORMANCE END DATE: 02/28/2009

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GENE EXPRESSION IN ENTAMOEBA HISTOLYTICA - DESCRIPTION (provided by the applicant): We propose to study gene expression in Entamoeba histolytica controlled by the upstream regulatory element 3 binding protein (URE3-BP). We hypothesize that URE3-BP regulates virulence by a coordinated regulation of.qene expression in response to changes in intracellular Ca 2*. This sequence-specific DNA binding protein recognizes the URE3 sequence present in the promoters of the Gal/GalNAc-inhibitible lectin hgl5 and ferredoxin 1 fdx genes. URE3-BP contains two Ca2*-binding EF-hand motifs, and increases in Ca 2v block its ability to bind to URE3 in vitro and to URE3-containing promoters in vivo. Mutation of the second EF hand motif in URE3-BP leads to a loss of Ca 2. inhibition of DNA binding. These results demonstrate that elevated intracellular Ca 2vblocks the ability of URE3-BP to bind to URE3-containing promoters. Modulation of URE3-BP by intracellular Ca 2v may represent an important mechanism of control of gene expression in E. histolytica. Three specific aims are proposed to test our hypothesis: (1) Study of the URE3-BP - URE3 cisacting DNA sequence interaction, including definition of the DNA motif required for the interaction as determined by experimental and computer-assisted modeling; (2) Study of the structure and function of URE3-BP, including mapping residues important in binding to calcium, DNA and membranes as well as domains of the molecule important for intracellular localization; and (3) Determination of gene expression controlled by URE3-BP, including Entamoeba genomic micro-array analysis of global gene expression and chromatin immunoprecipitation using anti-URE3-BP mAb. Successful completion of these studies will provide insight into transcription regulation of virulence factors by the novel mechanism of direct calcium control of an EF-hand containing transcription factor.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2008 ( Subtotal = $268,140 )
2008	2008	UNIVERSITY OF VIRGINIA	PO BOX 400195	CHARLOTTESVILLE	VA	22904	CHARLOTTESVILLE CITY	USA	Allergy and Infectious Diseases Research	000	13	2/29/2008	NON-COMPETING CONTINUATION	$268,140
														Subtotal = $268,140

Issue Date FY: 2007 ( Subtotal = $273,736 )
2007	2007	UNIVERSITY OF VIRGINIA	PO BOX 400195	CHARLOTTESVILLE	VA	22904	CHARLOTTESVILLE CITY	USA	Allergy and Infectious Diseases Research	000	12	2/28/2007	NON-COMPETING CONTINUATION	$273,736
														Subtotal = $273,736

Issue Date FY: 2006 ( Subtotal = $282,315 )
2006	2006	UNIVERSITY OF VIRGINIA	PO BOX 400195	CHARLOTTESVILLE	VA	22904	CHARLOTTESVILLE CITY	USA	Allergy and Infectious Diseases Research	000	11	2/6/2006	NON-COMPETING CONTINUATION	$282,315
														Subtotal = $282,315

Issue Date FY: 2005 ( Subtotal = $289,387 )
2005	2005	UNIVERSITY OF VIRGINIA	PO BOX 9003	CHARLOTTESVILLE	VA	22906	CHARLOTTESVILLE CITY	USA	Microbiology and Infectious Diseases Research	000	10	1/21/2005	NON-COMPETING CONTINUATION	$289,387
														Subtotal = $289,387

Issue Date FY: 2004 ( Subtotal = $301,883 )
2004	2004	UNIVERSITY OF VIRGINIA	PO BOX 9003	CHARLOTTESVILLE	VA	22906	CHARLOTTESVILLE CITY	USA	Microbiology and Infectious Diseases Research	000	9	2/29/2004	COMPETING CONTINUATION	$301,883
														Subtotal = $301,883

Issue Date FY: 2003 ( Subtotal = $229,378 )
2003	2003	UNIVERSITY OF VIRGINIA	PO BOX 9003	CHARLOTTESVILLE	VA	22906	CHARLOTTESVILLE CITY	USA	Microbiology and Infectious Diseases Research	000	8		NON-COMPETING CONTINUATION	$229,378
														Subtotal = $229,378

Issue Date FY: 2002 ( Subtotal = $224,000 )
2002	2002	UNIVERSITY OF VIRGINIA	PO BOX 9003	CHARLOTTESVILLE	VA	22906	CHARLOTTESVILLE CITY	USA	Microbiology and Infectious Diseases Research	000	7	1/24/2002	NON-COMPETING CONTINUATION	$224,000
														Subtotal = $224,000

Issue Date FY: 2001 ( Subtotal = $218,780 )
2001	2001	UNIVERSITY OF VIRGINIA	PO BOX 9003	CHARLOTTESVILLE	VA	22906	CHARLOTTESVILLE CITY	USA	Microbiology and Infectious Diseases Research	000	6	3/9/2001	NON-COMPETING CONTINUATION	$218,780
														Subtotal = $218,780

Issue Date FY: 2000 ( Subtotal = $234,248 )
2000	2000	UNIVERSITY OF VIRGINIA	PO BOX 400195	CHARLOTTESVILLE	VA	22904	CHARLOTTESVILLE CITY	USA	Microbiology and Infectious Diseases Research	000	5	3/24/2000	NON-COMPETING CONTINUATION	$213,712
2000	2000	UNIVERSITY OF VIRGINIA	PO BOX 400195	CHARLOTTESVILLE	VA	22904	CHARLOTTESVILLE CITY	USA	Microbiology and Infectious Diseases Research	001	5	7/12/2000	SUPPLEMENT FOR EXPANSION	$20,536
														Subtotal = $234,248

Issue Date FY: 1999 ( Subtotal = $207,691 )
1999	1999	UNIVERSITY OF VIRGINIA	PO BOX 400195	CHARLOTTESVILLE	VA	22904	CHARLOTTESVILLE CITY	USA	Microbiology and Infectious Diseases Research	000	4	2/15/1999	COMPETING CONTINUATION	$207,691
														Subtotal = $207,691

Issue Date FY: 1998 ( Subtotal = $177,012 )
1998	1998	UNIVERSITY OF VIRGINIA	PO BOX 400195	CHARLOTTESVILLE	VA	22904	CHARLOTTESVILLE CITY	USA	Microbiology and Infectious Diseases Research	000	3	1/7/1998	NON-COMPETING CONTINUATION	$177,012
														Subtotal = $177,012

Grand Total All Awards = $2,706,570

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GENE EXPRESSION IN ENTAMOEBA HISTOLYTICA

Award Number: R01AI037941

ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 02/01/1996

PERIOD OF PERFORMANCE END DATE: 02/28/2009

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