Preparation and characterization of soluble macronuclear chromatin from the hypotrich Euplotes eurystomus

Carmen L. Cadilla; Joel Harp; John M. Flanagan; Ada L. Olins; Donald E. Olins

doi:10.1093/nar/14.2.823

Preparation and characterization of soluble macronuclear chromatin from the hypotrich Euplotes eurystomus

Carmen L. Cadilla, Joel Harp, John M. Flanagan, Ada L. Olins, Donald E. Olins

Research output: Contribution to journal › Article › peer-review

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Abstract

Euplotes eurystomus la a hypotrichous ciliate containing a transcrip-tionally active nacronucleus (MAC) and a transcriptionally inactive micronucleus. Soluble MAC chromatin contains a normal complement of inner his tones, an Hl-like protein which is very sensitive to proteolysis, and a considerable proportion of non-histone proteins. A combination of N-Tosyl lysine chloromethyl ketone (TLCK) and PMSF was found to be most effective in preventing proteolysls. Mlcroccocal nuclease digestion yielded an average nucleosome repeat length of 187 ± 25 bp for soluble chroaatin; and 190 ± 9 bp for isolated nacronuclei. Thermal denaturatlon profiles of MAC chromatin in 0.25 nM EDTA display two main transitions at about 76 and 83°C, resembling the melting of soluble chicken erythrocyte chromatin. Circular dichroic spectra of MAC chromatin were compared to soluble chicken erythrocyte chromatin under the same ionic strength conditions and were found to be very similar. 20 chromatin/DNA agarose gel electrophoresis resulted in a diagonal line of DNA staining, which establishes a strict correlation between DNA and chromatin electrophoretic mobility.

Original language	English (US)
Pages (from-to)	823-841
Number of pages	19
Journal	Nucleic acids research
Volume	14
Issue number	2
DOIs	https://doi.org/10.1093/nar/14.2.823
State	Published - Jan 26 1986

All Science Journal Classification (ASJC) codes

Genetics

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@article{1669d2bc1f5745a28cd130c23ee25b59,

title = "Preparation and characterization of soluble macronuclear chromatin from the hypotrich Euplotes eurystomus",

abstract = "Euplotes eurystomus la a hypotrichous ciliate containing a transcrip-tionally active nacronucleus (MAC) and a transcriptionally inactive micronucleus. Soluble MAC chromatin contains a normal complement of inner his tones, an Hl-like protein which is very sensitive to proteolysis, and a considerable proportion of non-histone proteins. A combination of N-Tosyl lysine chloromethyl ketone (TLCK) and PMSF was found to be most effective in preventing proteolysls. Mlcroccocal nuclease digestion yielded an average nucleosome repeat length of 187 ± 25 bp for soluble chroaatin; and 190 ± 9 bp for isolated nacronuclei. Thermal denaturatlon profiles of MAC chromatin in 0.25 nM EDTA display two main transitions at about 76 and 83°C, resembling the melting of soluble chicken erythrocyte chromatin. Circular dichroic spectra of MAC chromatin were compared to soluble chicken erythrocyte chromatin under the same ionic strength conditions and were found to be very similar. 20 chromatin/DNA agarose gel electrophoresis resulted in a diagonal line of DNA staining, which establishes a strict correlation between DNA and chromatin electrophoretic mobility.",

author = "Cadilla, {Carmen L.} and Joel Harp and Flanagan, {John M.} and Olins, {Ada L.} and Olins, {Donald E.}",

note = "Funding Information: •Research supported by NIH research grants GM-19334 to D.E.O., and to A.L.O. (GM-29429), by an NIGMS National Research Service Award to C.L.C. (GM-10079-02) and by the Office of Health and Environmental Research, D.S. Depart»ent of Energy, under contract DE-AC05-840R21400 with the Martin Marietta Energy Systems, Inc. Funding Information: ACKNOWLEDGEMENTS We thank James Finch for excellent photographic work. This research was sponsored by the U.S. Department of Energy, under contract DE-AE05-84R2100 with the Martin Marietta Energy Systems, Inc., by grants from NIH to D.E.O. (GM-19334), and to A.L.O. (GM-29429), and by an NIGMS National Research Service Award to C.L.C. (GM-10079-02).",

year = "1986",

month = jan,

day = "26",

doi = "10.1093/nar/14.2.823",

language = "English (US)",

volume = "14",

pages = "823--841",

journal = "Nucleic acids research",

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T1 - Preparation and characterization of soluble macronuclear chromatin from the hypotrich Euplotes eurystomus

AU - Cadilla, Carmen L.

AU - Harp, Joel

AU - Flanagan, John M.

AU - Olins, Ada L.

AU - Olins, Donald E.

N1 - Funding Information: •Research supported by NIH research grants GM-19334 to D.E.O., and to A.L.O. (GM-29429), by an NIGMS National Research Service Award to C.L.C. (GM-10079-02) and by the Office of Health and Environmental Research, D.S. Depart»ent of Energy, under contract DE-AC05-840R21400 with the Martin Marietta Energy Systems, Inc. Funding Information: ACKNOWLEDGEMENTS We thank James Finch for excellent photographic work. This research was sponsored by the U.S. Department of Energy, under contract DE-AE05-84R2100 with the Martin Marietta Energy Systems, Inc., by grants from NIH to D.E.O. (GM-19334), and to A.L.O. (GM-29429), and by an NIGMS National Research Service Award to C.L.C. (GM-10079-02).

PY - 1986/1/26

Y1 - 1986/1/26

N2 - Euplotes eurystomus la a hypotrichous ciliate containing a transcrip-tionally active nacronucleus (MAC) and a transcriptionally inactive micronucleus. Soluble MAC chromatin contains a normal complement of inner his tones, an Hl-like protein which is very sensitive to proteolysis, and a considerable proportion of non-histone proteins. A combination of N-Tosyl lysine chloromethyl ketone (TLCK) and PMSF was found to be most effective in preventing proteolysls. Mlcroccocal nuclease digestion yielded an average nucleosome repeat length of 187 ± 25 bp for soluble chroaatin; and 190 ± 9 bp for isolated nacronuclei. Thermal denaturatlon profiles of MAC chromatin in 0.25 nM EDTA display two main transitions at about 76 and 83°C, resembling the melting of soluble chicken erythrocyte chromatin. Circular dichroic spectra of MAC chromatin were compared to soluble chicken erythrocyte chromatin under the same ionic strength conditions and were found to be very similar. 20 chromatin/DNA agarose gel electrophoresis resulted in a diagonal line of DNA staining, which establishes a strict correlation between DNA and chromatin electrophoretic mobility.

AB - Euplotes eurystomus la a hypotrichous ciliate containing a transcrip-tionally active nacronucleus (MAC) and a transcriptionally inactive micronucleus. Soluble MAC chromatin contains a normal complement of inner his tones, an Hl-like protein which is very sensitive to proteolysis, and a considerable proportion of non-histone proteins. A combination of N-Tosyl lysine chloromethyl ketone (TLCK) and PMSF was found to be most effective in preventing proteolysls. Mlcroccocal nuclease digestion yielded an average nucleosome repeat length of 187 ± 25 bp for soluble chroaatin; and 190 ± 9 bp for isolated nacronuclei. Thermal denaturatlon profiles of MAC chromatin in 0.25 nM EDTA display two main transitions at about 76 and 83°C, resembling the melting of soluble chicken erythrocyte chromatin. Circular dichroic spectra of MAC chromatin were compared to soluble chicken erythrocyte chromatin under the same ionic strength conditions and were found to be very similar. 20 chromatin/DNA agarose gel electrophoresis resulted in a diagonal line of DNA staining, which establishes a strict correlation between DNA and chromatin electrophoretic mobility.

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JO - Nucleic acids research

JF - Nucleic acids research

IS - 2

ER -

Preparation and characterization of soluble macronuclear chromatin from the hypotrich Euplotes eurystomus

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