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Structure of the macronuclear polyubiquitin gene in Euplotes

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Abstract

The hypotrichous ciliate, Euplotes eurystomus, contains both a transcriptionally inactive micronucleus (MIC) and a transcriptionally active macronucleus (MAC) in the same cell. MAC DNA is small (0.5–20 kb), linear and highly amplified. Each DNA fragment consists of two telomeres, a single coding region, and the necessary control elements to regulate gene transcription and replication. The polyubiquitin gene consists of 898 bp, plus 28 bp of double-stranded and 14 bases of single-stranded DNA of the telomeric repeat G4T4 at each end. The coding region exists as three copies of the ubiquitin gene (690 bp) fused in a head-to-tail arrangement as in other organisms. The stop codon is TAA, as in other Euplotes genes, and is not the rare glutamine codon used in most other ciliates. The 3′ nontranslated region contains two presumptive poly(A) addition sites; the 5′ nontranslated region possesses two putative TATA boxes, several imperfect direct and inverted repeats, and a possible origin of replication. Nucleosome positioning studies reveal four tightly packed nucleosomes and a non-nucleosomal area containing the probable 5′ control region as well as part of the coding region. The 5′ area does not contain any DNAse I hypersensitive sites. Although the telomeres are protected from exonuclease digestion, they are not as well protected as Oxytricha telomeres against endonucleases and cleavage by methidium propyl Fe+ EDTA.

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Abbreviations

MIC:

micronulceus

MAC:

macronucleus

MPE·Fe2+ :

methidium propyl EDTA·Fe2+

DNAse I:

deoxyribonuclease I

MNase:

micrococcal nuclease

RNAse A:

ribonuclease A

SDS:

sodium dodecyl sulfate

20×SSC, 3 M NaCl:

300 mM sodium citrate

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Authors and Affiliations

  1. Biology Division, Oak Ridge National Laboratory, U.T.-Oak Ridge Graduate School of Biomedical Sciences, P.O. Box 2009, 37831-8077, Oak Ridge, TN, USA

    Loren J. Hauser, Arthur E. Roberson & Donald E. Olins

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  1. Loren J. Hauser
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  2. Arthur E. Roberson
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  3. Donald E. Olins
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by D. Bazett-Jones

Research sponsored by the Office of Health and Environmental Research, United States Department of Energy, under Contract DE-AC05-840R21400 with the Martin Marietta Energy Systems, by NSF Research Grant DCB870057 to D.E.O, by NCI Training Grant T32CA09336, and by NIH Genetics Training Grant GM07438 to U.T.-Oak Ridge Graduate School of Biomedical Sciences

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Hauser, L.J., Roberson, A.E. & Olins, D.E. Structure of the macronuclear polyubiquitin gene in Euplotes . Chromosoma 100, 386–394 (1991). https://doi.org/10.1007/BF00337517

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