Our research analyzes molecular mechanisms regulating chorion (eggshell) gene expression in time and space within ovarian follicle cells during Drosophila oogenesis. The specific amplification of the two major chorion gene clusters remains a primary focus of study. We will further delineate cis-regulatory sequences required for amplification by analyzing control regions mutated in vitro via P element-mediated transformation. The role of transcriptional control elements, including enhancers, in defining the tissue-specificity of amplification will be tested. This will require further definition of the cis-regulatory elements of the s15, s18 and possibly other chorion genes. Replication origins used during amplification will be mapped to determine if they correspond to genetically defined essential sequences for amplification. We will attempt to develop a general method for mapping Drosophila replication origins using minichromosomes which can replicate extrachromosomally in Drosophila cells. We will analyze and clone unlinked genes which are required for chorion gene amplification, including fs(3)293 and fs(3)272. Additional genes in this class will be identified using a method of insertional mutagenesis by single P element hopping which greatly simplifies the task of cloning genetically defined Drosophila genes. Follicle cell nuclear proteins binding to amplification control regions will be purified and used to identify the corresponding genes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM027875-09
Application #
3275094
Study Section
Genetics Study Section (GEN)
Project Start
1980-04-01
Project End
1993-03-31
Budget Start
1988-04-01
Budget End
1989-03-31
Support Year
9
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Carnegie Institution of Washington, D.C.
Department
Type
DUNS #
072641707
City
Washington
State
DC
Country
United States
Zip Code
20005
Kotova, Elena; Jarnik, Michael; Tulin, Alexei V (2010) Uncoupling of the transactivation and transrepression functions of PARP1 protein. Proc Natl Acad Sci U S A 107:6406-11
Ji, Yingbiao; Tulin, Alexei V (2009) Poly(ADP-ribosyl)ation of heterogeneous nuclear ribonucleoproteins modulates splicing. Nucleic Acids Res 37:3501-13
Kotova, Elena; Jarnik, Michael; Tulin, Alexei V (2009) Poly (ADP-ribose) polymerase 1 is required for protein localization to Cajal body. PLoS Genet 5:e1000387
Pinnola, Aaron; Naumova, Natasha; Shah, Meera et al. (2007) Nucleosomal core histones mediate dynamic regulation of poly(ADP-ribose) polymerase 1 protein binding to chromatin and induction of its enzymatic activity. J Biol Chem 282:32511-9
Tulin, Alexei; Chinenov, Yurii; Spradling, Allan (2003) Regulation of chromatin structure and gene activity by poly(ADP-ribose) polymerases. Curr Top Dev Biol 56:55-83
Tulin, Alexei; Spradling, Allan (2003) Chromatin loosening by poly(ADP)-ribose polymerase (PARP) at Drosophila puff loci. Science 299:560-2
Tulin, Alexei; Stewart, Dianne; Spradling, Allan C (2002) The Drosophila heterochromatic gene encoding poly(ADP-ribose) polymerase (PARP) is required to modulate chromatin structure during development. Genes Dev 16:2108-19
Calvi, B R; Spradling, A C (2001) The nuclear location and chromatin organization of active chorion amplification origins. Chromosoma 110:159-72
Lilly, M A; de Cuevas, M; Spradling, A C (2000) Cyclin A associates with the fusome during germline cyst formation in the Drosophila ovary. Dev Biol 218:53-63
Dej, K J; Spradling, A C (1999) The endocycle controls nurse cell polytene chromosome structure during Drosophila oogenesis. Development 126:293-303

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