Cloning of Abundant MRNA Species Present During Conjugation of Tetrahymena Thermophila: Identification of MRNA Species Present Exclusively During Meiosis

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 1983 Oct 1

PMID 6646127

Citations 32

Authors

D W Martindale

P J Bruns

Affiliations

Soon will be listed here.

Abstract

A cDNA library was constructed by using as a template the RNA present during the meiotic prophase of Tetrahymena thermophila. Clones containing cDNA sequences homologous to moderately abundant to abundant transcripts were detected by colony hybridization and confirmed by hybridizing purified cDNA plasmids on filters with labeled RNA probes. Eighteen clones were isolated, and the sizes of their cDNA inserts were determined. Cross-hybridization of individual cDNA plasmid pairs showed that each of these clones contained cDNA that was homologous to one of eight different RNA transcripts. The sizes of the eight RNA transcripts and the stages of the T. thermophila life cycle during which they were present were determined by hybridizing nick-translated cDNA probes against denatured, electrophoresed RNA from various stages. Clones were identified that contained sequences homologous to RNAs present only in early conjugation (meiosis); other clones contained sequences homologous to RNAs which were abundant during conjugation but present at other stages as well. One clone contained a sequence homologous to an RNA that was abundantly present only in nongrowing cells.

Citing Articles

Functional Proteomics of Nuclear Proteins in A Review.

Saettone A, Nabeel-Shah S, Garg J, Lambert J, Pearlman R, Fillingham J Genes (Basel). 2019; 10(5).

PMID: 31052454 PMC: 6562869. DOI: 10.3390/genes10050333.

Two GW repeat proteins interact with Tetrahymena thermophila argonaute and promote genome rearrangement.

Bednenko J, Noto T, Desouza L, Siu K, Pearlman R, Mochizuki K Mol Cell Biol. 2009; 29(18):5020-30.

PMID: 19596782 PMC: 2738283. DOI: 10.1128/MCB.00076-09.

Refined annotation and assembly of the Tetrahymena thermophila genome sequence through EST analysis, comparative genomic hybridization, and targeted gap closure.

Coyne R, Thiagarajan M, Jones K, Wortman J, Tallon L, Haas B BMC Genomics. 2008; 9:562.

PMID: 19036158 PMC: 2612030. DOI: 10.1186/1471-2164-9-562.

Isolation and characterization of the actin gene from Tetrahymena thermophila.

Cupples C, Pearlman R Proc Natl Acad Sci U S A. 1986; 83(14):5160-4.

PMID: 16593729 PMC: 323910. DOI: 10.1073/pnas.83.14.5160.

The H1 phosphorylation state regulates expression of CDC2 and other genes in response to starvation in Tetrahymena thermophila.

Dou Y, Song X, Liu Y, Gorovsky M Mol Cell Biol. 2005; 25(10):3914-22.

PMID: 15870266 PMC: 1087734. DOI: 10.1128/MCB.25.10.3914-3922.2005.

References

Wahl G, Padgett R, Stark G . Gene amplification causes overproduction of the first three enzymes of UMP synthesis in N-(phosphonacetyl)-L-aspartate-resistant hamster cells. J Biol Chem. 1979; 254(17):8679-89. View

Biessmann H, Craig E, McCarthy B . Rapid quantitation of individual RNA species in a complex population. Nucleic Acids Res. 1979; 7(4):981-96. PMC: 342276. DOI: 10.1093/nar/7.4.981. View

Spradling A, Digan M, Mahowald A, Scott M, Craig E . Two clusters of genes for major chorion proteins of Drosophila melanogaster. Cell. 1980; 19(4):905-14. DOI: 10.1016/0092-8674(80)90082-3. View

Hanahan D, Meselson M . Plasmid screening at high colony density. Gene. 1980; 10(1):63-7. DOI: 10.1016/0378-1119(80)90144-4. View

Thomas P . Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980; 77(9):5201-5. PMC: 350025. DOI: 10.1073/pnas.77.9.5201. View

Holmes D, Quigley M . A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981; 114(1):193-7. DOI: 10.1016/0003-2697(81)90473-5. View

Martindale D, Allis C, Bruns P . Conjugation in Tetrahymena thermophila. A temporal analysis of cytological stages. Exp Cell Res. 1982; 140(1):227-36. DOI: 10.1016/0014-4827(82)90172-0. View

Bleyman L, Simon E . Genetic control of maturity in Tetrahymena pyriformis. Genet Res. 1967; 10(3):319-21. DOI: 10.1017/s0016672300011083. View

Boyer H . A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969; 41(3):459-72. DOI: 10.1016/0022-2836(69)90288-5. View

10.

Aviv H, Leder P . Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972; 69(6):1408-12. PMC: 426713. DOI: 10.1073/pnas.69.6.1408. View

11.

GODSON G, Vapnek D . A simple method of preparing large amounts of phiX174 RF 1 supercoiled DNA. Biochim Biophys Acta. 1973; 299(4):516-20. DOI: 10.1016/0005-2787(73)90223-2. View

12.

Bruns P, Brussard T . Pair formation in tetrahymena pyriformis, an inducible developmental system. J Exp Zool. 1974; 188(3):337-44. DOI: 10.1002/jez.1401880309. View

13.

Sugai T, Hiwatashi K . Cytologic and autoradiographic studies of the micronucleus at meiotic prophase in Tetrahymena pyriformis. J Protozool. 1974; 21(4):542-8. DOI: 10.1111/j.1550-7408.1974.tb03695.x. View

14.

Southern E . Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975; 98(3):503-17. DOI: 10.1016/s0022-2836(75)80083-0. View

15.

Grunstein M, Hogness D . Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975; 72(10):3961-5. PMC: 433117. DOI: 10.1073/pnas.72.10.3961. View

16.

Maizels N . Dictyostelium 17S, 25S, and 5S rDNAs lie within a 38,000 base pair repeated unit. Cell. 1976; 9(3):431-8. DOI: 10.1016/0092-8674(76)90088-x. View

17.

Rigby P, Dieckmann M, Rhodes C, Berg P . Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977; 113(1):237-51. DOI: 10.1016/0022-2836(77)90052-3. View

18.

McMaster G, Carmichael G . Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977; 74(11):4835-8. PMC: 432050. DOI: 10.1073/pnas.74.11.4835. View

19.

Bolivar F, Rodriguez R, Greene P, Betlach M, Heyneker H, Boyer H . Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977; 2(2):95-113. View

20.

Efstratiadis A, Broome S, Lomedico P, Tizard R, Naber S, Chick W . A bacterial clone synthesizing proinsulin. Proc Natl Acad Sci U S A. 1978; 75(8):3727-31. PMC: 392859. DOI: 10.1073/pnas.75.8.3727. View