The proposed studies deal with nuclear proteins and their assembly into macromolecular complexes with RNA on the chromosomes and in other nuclear organelles. Advantage will be taken of the giant nucleus and giant lampbrush chromosomes (LBCs) of the amphibian oocyte. Monoclonal antibodies will be produced against nuclear proteins, whose subnuclear distribution will be determined by immunofluorescence. The extraordinary morphological detail in LBCs allows antigen detection within single transcription units or in defined subnuclear particulates. The antibodies will be used to recover cDNA clones from expression libraries. Protein sequences derived from the cDNA clones will permit deductions about the role(s) of the proteins in nuclear function. In vitro transcriptions made from the cDNA clones will be injected into amphibian oocytes, and the translation products (polypeptides) will be followed biochemically and cytologically from their site of synthesis in the cytoplasm through the nuclear envelope to their final destination on the chromosomes or elsewhere in the nucleus. By appropriate in vitro alteration of the injected transcripts we will determine what amino acid sequences are necessary for transport across the nuclear envelope and for targeting to the final subnuclear destination. Studies will also be conducted on the transcription of the 5S gene sequences on LBCs by in situ nucleic acid hybridization. Finally studies will be carried out on a novel selfcleaving RNA transcribed from a short, repeated sequence of newt DNA, with emphasis on the possible existence of a ribonucleoprotein particle. All of the proposed studies deal with how RNA produced by the chromosomes becomes associated with nuclear proteins in order to carry out basic cellular functions (RNA processing, storage, and transport). Knowledge of such functions is fundamental for understanding cell metabolism in both the normal and diseased state.
| Deryusheva, Svetlana; Choleza, Maria; Barbarossa, Adrien et al. (2012) Post-transcriptional modification of spliceosomal RNAs is normal in SMN-deficient cells. RNA 18:31-6 |
| Singer, Alison B; Gall, Joseph G (2011) An inducible nuclear body in the Drosophila germinal vesicle. Nucleus 2:403-9 |
| Nizami, Zehra; Deryusheva, Svetlana; Gall, Joseph G (2010) The Cajal body and histone locus body. Cold Spring Harb Perspect Biol 2:a000653 |
| Nizami, Z F; Deryusheva, S; Gall, J G (2010) Cajal bodies and histone locus bodies in Drosophila and Xenopus. Cold Spring Harb Symp Quant Biol 75:313-20 |
| Gall, Joseph G; Wu, Zheng'an (2010) Examining the contents of isolated Xenopus germinal vesicles. Methods 51:45-51 |
| Deryusheva, Svetlana; Gall, Joseph G (2009) Small Cajal body-specific RNAs of Drosophila function in the absence of Cajal bodies. Mol Biol Cell 20:5250-9 |
| Liu, Ji-Long; Wu, Zheng'an; Nizami, Zehra et al. (2009) Coilin is essential for Cajal body organization in Drosophila melanogaster. Mol Biol Cell 20:1661-70 |
| Gall, Joseph G (2009) Chromosome odds and ends. Annu Rev Cell Dev Biol 25:1-19 |
| Liu, Ji-Long; Gall, Joseph G (2007) U bodies are cytoplasmic structures that contain uridine-rich small nuclear ribonucleoproteins and associate with P bodies. Proc Natl Acad Sci U S A 104:11655-9 |
| Handwerger, Korie E; Gall, Joseph G (2006) Subnuclear organelles: new insights into form and function. Trends Cell Biol 16:19-26 |
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