Differential Gene Expression During Stationary Phase Between Amicronucleates and Micronucleates of the Ciliated Protist, Pseudourostyla Cristata

Overview

Journal Curr Genet

Specialty Genetics

Date 2005 Nov 12

PMID 16283314

Authors

Xing-Yin Liu

Kam-Len Daniel Lee

Yong-Zhen Mao

Tao Liu

Li-Pei Jin

Affiliations

Soon will be listed here.

Abstract

Ciliates are unicellular eukaryotic organisms with two types of nuclei, the 'germline' micronucleus (MIC) and the 'somatic' macronucleus (MAC). We previously reported that when the MIC of Pseudourostyla cristata was eliminated by amputation, the resultant amicronucleate organisms exhibited a lower viability and abnormal oral structures. To gain insight into the genetic reorganization involved in or induced by removal of the MIC and the mechanism by which nuclear dimorphism was established, we investigated gene expression differences between amicronucleates and micronucleates, using suppression subtractive hybridization (SSH) techniques. Approximately 250 clones from each library were screened by cDNA array dot blotting. Altogether, 22 unique genes from the forward-subtractive library (micronucleates as tester, amicronucleates as driver) and 23 unique genes from the reverse-subtractive library (micronucleates as driver and amicronucleates as tester) were shown to be differentially expressed. These 45 differentially expressed genes were found to be homologs of genes involved in various cellular processes including signal transduction, transcription, cell cycle accomplishment and general metabolism, cell structure, and stress response. We highlighted 14 genes, 7 that were unique from both the forward-subtractive and the reverse-subtractive libraries, using real time semi-quantitative RT-PCR. The characterization of these cDNAs represents a starting point in understanding the molecular mechanisms of amicronucleates disruption.

References

Haremaki T, Sugai T, Takahashi M . Involvement of active cellular mechanisms on the disorganization of oral apparatus in amicronucleate cells in Tetrahymena thermophila. Cell Struct Funct. 1996; 21(1):73-80. DOI: 10.1247/csf.21.73. View

Liu X, Kim C, Yang J, Jemmerson R, Wang X . Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c. Cell. 1996; 86(1):147-57. DOI: 10.1016/s0092-8674(00)80085-9. View

Fetzer C, Hogan D, Lipps H . A PIWI homolog is one of the proteins expressed exclusively during macronuclear development in the ciliate Stylonychia lemnae. Nucleic Acids Res. 2002; 30(20):4380-6. PMC: 137143. DOI: 10.1093/nar/gkf579. View

Villalobo E, Moch C, Fryd-Versavel G, Fleury-Aubusson A, Morin L . Cysteine proteases and cell differentiation: excystment of the ciliated protist Sterkiella histriomuscorum. Eukaryot Cell. 2003; 2(6):1234-45. PMC: 326638. DOI: 10.1128/EC.2.6.1234-1245.2003. View

Martinou J, Desagher S, Antonsson B . Cytochrome c release from mitochondria: all or nothing. Nat Cell Biol. 2000; 2(3):E41-3. DOI: 10.1038/35004069. View

Haremaki T, Sugai T, Takahashi M . The vegetative micronucleus has a critical role in maintenance of cortical structure in Tetrahymena thermophila. Cell Struct Funct. 1995; 20(3):239-44. DOI: 10.1247/csf.20.239. View

Moreira M, del Portillo H, Milder R, Balanco J, Barcinski M . Heat shock induction of apoptosis in promastigotes of the unicellular organism Leishmania (Leishmania) amazonensis. J Cell Physiol. 1996; 167(2):305-13. DOI: 10.1002/(SICI)1097-4652(199605)167:2<305::AID-JCP15>3.0.CO;2-6. View

Lu E, Wolfe J . Lysosomal enzymes in the macronucleus of Tetrahymena during its apoptosis-like degradation. Cell Death Differ. 2001; 8(3):289-97. DOI: 10.1038/sj.cdd.4400807. View

Welburn S, Murphy N . Prohibitin and RACK homologues are up-regulated in trypanosomes induced to undergo apoptosis and in naturally occurring terminally differentiated forms. Cell Death Differ. 1999; 5(7):615-22. DOI: 10.1038/sj.cdd.4400393. View

10.

Kirk D . Germ cell specification in Volvox carteri. Ciba Found Symp. 1994; 182:2-15; discussion 15-30. DOI: 10.1002/9780470514573.ch2. View

11.

Noguchi M, Kurahashi S, Kamachi H, Inoue H . Control of the ciliary beat by cyclic nucleotides in intact cortical sheets from Paramecium. Zoolog Sci. 2004; 21(12):1167-75. DOI: 10.2108/zsj.21.1167. View

12.

Plattner H, Klauke N . Calcium in ciliated protozoa: sources, regulation, and calcium-regulated cell functions. Int Rev Cytol. 2000; 201:115-208. DOI: 10.1016/s0074-7696(01)01003-8. View

13.

Mikami K . Stomatogenesis during sexual and asexual reproduction in an amicronucleate strain of Paramecium caudatum. J Exp Zool. 1979; 208(1):121-8. DOI: 10.1002/jez.1402080113. View

14.

Yang T, Perasso R, Baroin-Tourancheau A . Myb genes in ciliates: a common origin with the myb protooncogene?. Protist. 2003; 154(2):229-38. DOI: 10.1078/143446103322166527. View

15.

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

16.

Grobe K, Poppelmann M, Becker W, Petersen A . Properties of group I allergens from grass pollen and their relation to cathepsin B, a member of the C1 family of cysteine proteinases. Eur J Biochem. 2002; 269(8):2083-92. DOI: 10.1046/j.1432-1033.2002.02856.x. View

17.

Simm R, Morr M, Kader A, Nimtz M, Romling U . GGDEF and EAL domains inversely regulate cyclic di-GMP levels and transition from sessility to motility. Mol Microbiol. 2004; 53(4):1123-34. DOI: 10.1111/j.1365-2958.2004.04206.x. View

18.

Prescott D . The DNA of ciliated protozoa. Microbiol Rev. 1994; 58(2):233-67. PMC: 372963. DOI: 10.1128/mr.58.2.233-267.1994. View

19.

Shi X, Frankel J . Morphology and development of mirror-image doublets of Stylonychia mytilus. J Protozool. 1990; 37(1):1-13. DOI: 10.1111/j.1550-7408.1990.tb01106.x. View

20.

Diatchenko L, Lukyanov S, Lau Y, Siebert P . Suppression subtractive hybridization: a versatile method for identifying differentially expressed genes. Methods Enzymol. 1999; 303:349-80. DOI: 10.1016/s0076-6879(99)03022-0. View