The long-term goal of this project is to determine how a signal originating at the cell periphery triggers a series of events culminating in the synthesis and assembly of an intracellular organelle. Specifically, we are studying the mechanisms by which deciliation of the ciliated protozoan Tetrahymena thermophila initiates the orderly expression of specific genes required for reformation of cilia. Improved methods have been developed for obtaining viable deciliation of Tetrahymena which induces a rapid (approximately l0-fold) induction of protein synthesis and a specific, relative (approximately l0-fold) co-ordinate induction of alpha and beta tubulins. Translational and transcriptional mechanisms underlying this general induction of protein synthesis and specific induction of tubulin are being examined. Polysomal polyA ion RNA isolated from regenerating cells 80-l00 min. after deciliation (at which time tubulin represents approximately l0% of total protein synthesis) is being utilized as a source of cDNA from which tubulin gene sequences will be prepared by recombinant DNA techniques. These cloned DNA sequences will then be utilized to study the organization of tubulin genes and their expression during regeneration.

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National Institute of General Medical Sciences (NIGMS)
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Molecular Biology Study Section (MBY)
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University of Rochester
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Wloga, Dorota; Rogowski, Krzysztof; Sharma, Neeraj et al. (2008) Glutamylation on alpha-tubulin is not essential but affects the assembly and functions of a subset of microtubules in Tetrahymena thermophila. Eukaryot Cell 7:1362-72
Tsao, Che-Chia; Gorovsky, Martin A (2008) Tetrahymena IFT122A is not essential for cilia assembly but plays a role in returning IFT proteins from the ciliary tip to the cell body. J Cell Sci 121:428-36
Tsao, Che-Chia; Gorovsky, Martin A (2008) Different effects of Tetrahymena IFT172 domains on anterograde and retrograde intraflagellar transport. Mol Biol Cell 19:1450-61
Xie, Rong; Clark, Kathleen M; Gorovsky, Martin A (2007) Endoplasmic reticulum retention signal-dependent glycylation of the Hsp70/Grp170-related Pgp1p in Tetrahymena. Eukaryot Cell 6:388-97
Williams, Norman E; Tsao, Che-Chia; Bowen, Josephine et al. (2006) The actin gene ACT1 is required for phagocytosis, motility, and cell separation of Tetrahymena thermophila. Eukaryot Cell 5:555-67
Shang, Yuhua; Tsao, Che-Chia; Gorovsky, Martin A (2005) Mutational analyses reveal a novel function of the nucleotide-binding domain of gamma-tubulin in the regulation of basal body biogenesis. J Cell Biol 171:1035-44
Thazhath, Rupal; Jerka-Dziadosz, Maria; Duan, Jianming et al. (2004) Cell context-specific effects of the beta-tubulin glycylation domain on assembly and size of microtubular organelles. Mol Biol Cell 15:4136-47
Shang, Yuhua; Song, Xiaoyuan; Bowen, Josephine et al. (2002) A robust inducible-repressible promoter greatly facilitates gene knockouts, conditional expression, and overexpression of homologous and heterologous genes in Tetrahymena thermophila. Proc Natl Acad Sci U S A 99:3734-9
Shang, Yuhua; Li, Bing; Gorovsky, Martin A (2002) Tetrahymena thermophila contains a conventional gamma-tubulin that is differentially required for the maintenance of different microtubule-organizing centers. J Cell Biol 158:1195-206
Hai, B; Gaertig, J; Gorovsky, M A (2000) Knockout heterokaryons enable facile mutagenic analysis of essential genes in Tetrahymena. Methods Cell Biol 62:513-31

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