Abstract
The gene encoding the B type variable surface protein from Paramecium tetraurelia stock 51 has been cloned and sequenced. The 7,182 nucleotide open reading frame contains no introns and encodes a cysteine-rich protein that has a periodic structure including three nearly perfect tandem repeats in the central region. Interestingly, the B gene is located near a macronuclear telomere as was shown previously for two other paramecium surface protein genes. In this paper, we characterize four independent mutants with complete macronuclear deletions of the B gene. Previous analysis of different macronuclear deletion mutants of the A surface protein gene demonstrated two types of inheritance: typical Mendelian segregation (as illustrated by d12) and cytoplasmic inheritance (shown by d48). F(1) analysis of four B(-) mutants crossed with wild-type cells reveals heterozygous F(1) cell lines derived from both parental cytoplasms contain approximately the same copy number of the B gene, as expected for a recessive Mendelian mutation. Analysis of F(2) progeny from three of these four B(-) mutant crosses indicates that one of the three exhibits a Mendelian 1:1 segregation ratio of B(+) and B(-) cell lines. The other two show a preponderance of B(+) cells, but this is not correlated with the parental cytoplasmic type. In addition to having a large number of B(+) individuals, the d12.144, A(-), B(-) mutant produced some F(2) progeny that stably maintain less than normal macronuclear amounts of the A gene and/or the B gene.
Full Text
The Full Text of this article is available as a PDF (6.4 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
- Blackburn E. H., Karrer K. M. Genomic reorganization in ciliated protozoans. Annu Rev Genet. 1986;20:501–521. doi: 10.1146/annurev.ge.20.120186.002441. [DOI] [PubMed] [Google Scholar]
- Caron F. A high degree of macronuclear chromosome polymorphism is generated by variable DNA rearrangements in Paramecium primaurelia during macronuclear differentiation. J Mol Biol. 1992 Jun 5;225(3):661–678. doi: 10.1016/0022-2836(92)90393-x. [DOI] [PubMed] [Google Scholar]
- Caron F., Meyer E. Molecular basis of surface antigen variation in paramecia. Annu Rev Microbiol. 1989;43:23–42. doi: 10.1146/annurev.mi.43.100189.000323. [DOI] [PubMed] [Google Scholar]
- Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Epstein L. M., Forney J. D. Mendelian and non-mendelian mutations affecting surface antigen expression in Paramecium tetraurelia. Mol Cell Biol. 1984 Aug;4(8):1583–1590. doi: 10.1128/mcb.4.8.1583. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Forney J. D., Blackburn E. H. Developmentally controlled telomere addition in wild-type and mutant paramecia. Mol Cell Biol. 1988 Jan;8(1):251–258. doi: 10.1128/mcb.8.1.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Forney J. D., Epstein L. M., Preer L. B., Rudman B. M., Widmayer D. J., Klein W. H., Preer J. R., Jr Structure and expression of genes for surface proteins in Paramecium. Mol Cell Biol. 1983 Mar;3(3):466–474. doi: 10.1128/mcb.3.3.466. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Godiska R., Aufderheide K. J., Gilley D., Hendrie P., Fitzwater T., Preer L. B., Polisky B., Preer J. R., Jr Transformation of Paramecium by microinjection of a cloned serotype gene. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7590–7594. doi: 10.1073/pnas.84.21.7590. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Godiska R. Structure and sequence of the H surface protein gene of Paramecium and comparison with related genes. Mol Gen Genet. 1987 Jul;208(3):529–536. doi: 10.1007/BF00328151. [DOI] [PubMed] [Google Scholar]
- Harumoto T. Induced change in a non-mendelian determinant by transplantation of macronucleoplasm in Paramecium tetraurelia. Mol Cell Biol. 1986 Oct;6(10):3498–3501. doi: 10.1128/mcb.6.10.3498. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Henikoff S. Unidirectional digestion with exonuclease III in DNA sequence analysis. Methods Enzymol. 1987;155:156–165. doi: 10.1016/0076-6879(87)55014-5. [DOI] [PubMed] [Google Scholar]
- Jessop-Murray H., Martin L. D., Gilley D., Preer J. R., Jr, Polisky B. Permanent rescue of a non-Mendelian mutation of Paramecium by microinjection of specific DNA sequences. Genetics. 1991 Nov;129(3):727–734. doi: 10.1093/genetics/129.3.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koizumi S., Kobayashi S. Microinjection of plasmid DNA encoding the A surface antigen of Paramecium tetraurelia restores the ability to regenerate a wild-type macronucleus. Mol Cell Biol. 1989 Oct;9(10):4398–4401. doi: 10.1128/mcb.9.10.4398. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meyer E., Caron F., Baroin A. Macronuclear structure of the G surface antigen gene of Paramecium primaurelia and direct expression of its repeated epitopes in Escherichia coli. Mol Cell Biol. 1985 Sep;5(9):2414–2422. doi: 10.1128/mcb.5.9.2414. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nielsen E., You Y., Forney J. Cysteine residue periodicity is a conserved structural feature of variable surface proteins from Paramecium tetraurelia. J Mol Biol. 1991 Dec 20;222(4):835–841. doi: 10.1016/0022-2836(91)90573-o. [DOI] [PubMed] [Google Scholar]
- Prat A. Conserved sequences flank variable tandem repeats in two alleles of the G surface protein of Paramecium primaurelia. J Mol Biol. 1990 Feb 5;211(3):521–535. doi: 10.1016/0022-2836(90)90263-L. [DOI] [PubMed] [Google Scholar]
- Prat A., Katinka M., Caron F., Meyer E. Nucleotide sequence of the Paramecium primaurelia G surface protein. A huge protein with a highly periodic structure. J Mol Biol. 1986 May 5;189(1):47–60. doi: 10.1016/0022-2836(86)90380-3. [DOI] [PubMed] [Google Scholar]
- Preer J R. Studies on the Immobilization Antigens of Paramecium. IV. Properties of the Different Antigens. Genetics. 1959 Sep;44(5):803–814. doi: 10.1093/genetics/44.5.803. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Preer J. R., Jr, Preer L. B., Rudman B., Barnett A. Molecular biology of the genes for immobilization antigens in Paramecium. J Protozool. 1987 Nov;34(4):418–423. doi: 10.1111/j.1550-7408.1987.tb03205.x. [DOI] [PubMed] [Google Scholar]
- Preer J. R., Preer L. B., Rudman B. M. mRNAs for the immobilization antigens of Paramecium. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6776–6778. doi: 10.1073/pnas.78.11.6776. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reisner A. H., Rowe J., Sleigh R. W. Concerning the tertiary structure of the soluble surface proteins of Paramecium. Biochemistry. 1969 Nov;8(11):4637–4644. doi: 10.1021/bi00839a061. [DOI] [PubMed] [Google Scholar]
- Rudman B., Preer L. B., Polisky B., Preer J. R., Jr Mutants affecting processing of DNA in macronuclear development in paramecium. Genetics. 1991 Sep;129(1):47–56. doi: 10.1093/genetics/129.1.47. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- You Y., Aufderheide K., Morand J., Rodkey K., Forney J. Macronuclear transformation with specific DNA fragments controls the content of the new macronuclear genome in Paramecium tetraurelia. Mol Cell Biol. 1991 Feb;11(2):1133–1137. doi: 10.1128/mcb.11.2.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]