Ro ribonucleoprotein (Ro RNP) is the target of an aggressive autoimmune response in many patients with systemic lupus erythematosus (SLE) and Sjogren's syndrome (SS). Limited specificity of autoimmune responses in these disorders is consistent with the Ro RNP being important in the pathology and/or etiology of the autoimmune dysregulation found in these diseases. The function of Ro RNP is not known, though some structural data are available such as the sequences for the human (h) Y RNAs and the associated 60 kD Ro peptide. However, neither the secondary structures of the Y RNAs nor the conserved primary regions of Y RNA sequence between species have been established. Such data would have potential to reveal Ro RNP function, as was the case for U particle function in hnRNA splicing. The investigators propose to clone, sequence, and analyze the known Y RNAs from mice, rats, rabbits, cattle, ducks, and trout (16 Y RNAs). Also, toads, drosophila, lobster, a nematode, and yeast will be explored for the presence of Y-like RNAs and Ro-like mRNA. Finally, in order to define sufficient diversity of h Y5-like sequences it may be necessary to recover homologs of h Y5 from primates in addition to man. Technology to be used has been recently developed in the investigators' laboratory for these low abundance RNAs and with which hY4 RNA has been sequenced. Immunoprecipitated Y RNA will be polyadenylated followed by cDNA synthesis. After poly-g tailing, intervening cDNA or Y RNA will be expanded by polymerase chain reaction, cloned, and sequenced. With sequences categorized according to each type of Y RNA obtained, the preserved base pairing pattern will be used to construct the evolutionarily conserved secondary structure. Conserved primary sequence of homologous Y RNAs will provide short sequences with which to evaluate the existing DNA sequence data bases in an effort to find a pattern or recurring complementary sequences with potential functional significance. Such a result would lead toward elucidating a role for Ro RNPs in the molecular metabolism of cells. In the final phase of this project the investigators will develop a model system in which to generate and test hypotheses of Ro RNP function. Anti-sense DNA oligonucleotides prepared according to the 60 kD Ro and hY sequences will be evaluated for their capacity to inhibit Ro RNP formation. If Ro RNP formation is inhibited then the function of Ro RNP will be characterized in this model. If not, then the suitability of yeast as a model will be evaluated. If neither the anti-sense DNA oligos nor yeast provide suitable models, the investigators will transfect tissue culture cells with plasmid constructs that produce intracellular anti-sense RNAs designed to inhibit the products of the 60 kD Ro and hY genes. These experiments they believe will provide important structural and functional information that should carry them toward a comprehensive understanding of the Ro RNP antigen.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI031584-01
Application #
3146614
Study Section
General Medicine A Subcommittee 2 (GMA)
Project Start
1991-07-01
Project End
1994-06-30
Budget Start
1991-07-01
Budget End
1992-06-30
Support Year
1
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Oklahoma Medical Research Foundation
Department
Type
DUNS #
937727907
City
Oklahoma City
State
OK
Country
United States
Zip Code
73104
Harley, John B; Chen, Xiaoting; Pujato, Mario et al. (2018) Transcription factors operate across disease loci, with EBNA2 implicated in autoimmunity. Nat Genet 50:699-707
Liu, Ke; Kurien, Biji T; Zimmerman, Sarah L et al. (2016) X Chromosome Dose and Sex Bias in Autoimmune Diseases: Increased Prevalence of 47,XXX in Systemic Lupus Erythematosus and Sjögren's Syndrome. Arthritis Rheumatol 68:1290-1300
Kottyan, Leah C; Zoller, Erin E; Bene, Jessica et al. (2015) The IRF5-TNPO3 association with systemic lupus erythematosus has two components that other autoimmune disorders variably share. Hum Mol Genet 24:582-96
Morris, D L; Fernando, M M A; Taylor, K E et al. (2014) MHC associations with clinical and autoantibody manifestations in European SLE. Genes Immun 15:210-7
Molineros, Julio E; Maiti, Amit K; Sun, Celi et al. (2013) Admixture mapping in lupus identifies multiple functional variants within IFIH1 associated with apoptosis, inflammation, and autoantibody production. PLoS Genet 9:e1003222
Dozmorov, Igor; Dominguez, Nicolas; Sestak, Andrea L et al. (2013) Evidence of dynamically dysregulated gene expression pathways in hyperresponsive B cells from African American lupus patients. PLoS One 8:e71397
Vaughn, Samuel E; Kottyan, Leah C; Munroe, Melissa E et al. (2012) Genetic susceptibility to lupus: the biological basis of genetic risk found in B cell signaling pathways. J Leukoc Biol 92:577-91
Hughes, Travis; Adler, Adam; Merrill, Joan T et al. (2012) Analysis of autosomal genes reveals gene-sex interactions and higher total genetic risk in men with systemic lupus erythematosus. Ann Rheum Dis 71:694-9
Castillejo-López, Casimiro; Delgado-Vega, Angélica M; Wojcik, Jerome et al. (2012) Genetic and physical interaction of the B-cell systemic lupus erythematosus-associated genes BANK1 and BLK. Ann Rheum Dis 71:136-42
Bruner, Benjamin F; Guthridge, Joel M; Lu, Rufei et al. (2012) Comparison of autoantibody specificities between traditional and bead-based assays in a large, diverse collection of patients with systemic lupus erythematosus and family members. Arthritis Rheum 64:3677-86

Showing the most recent 10 out of 110 publications