Reconstitution of RNAase P activity using inactive subunits from E. coli and HeLa cells

Heidi A. Gold; Sidney Altman

doi:10.1016/0092-8674(86)90758-0

Reconstitution of RNAase P activity using inactive subunits from E. coli and HeLa cells

Heidi A. Gold, Sidney Altman

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

29 Scopus citations

Abstract

HeLa cell RNAase P activity found in the flow-through of anti-Sm affinity columns can be separated into inactive RNA and protein components. These components can be used to reconstitute active hybrid enzyme complexes with purified subunits from E. coli RNAase P. The RNA in the HeLa cell fractions employed is enriched for species between 85 and 115 nucleotides long. This reconstitution assay is a convenient means of purifying the functional RNA and protein of HeLa cell RNAase P. Probes derived from the genes for the subunits of E. coli RNAase P hybridize to genomic DNA of gram-negative prokaryotic organisms, but no positive signals are seen with genomic DNA from a variety of eukaryotic organisms.

Original language	English (US)
Pages (from-to)	243-249
Number of pages	7
Journal	Cell
Volume	44
Issue number	2
DOIs	https://doi.org/10.1016/0092-8674(86)90758-0
State	Published - Jan 31 1986
Externally published	Yes

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)

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10.1016/0092-8674(86)90758-0

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@article{ef05ced528da4f95ad770d312bfa4909,

title = "Reconstitution of RNAase P activity using inactive subunits from E. coli and HeLa cells",

abstract = "HeLa cell RNAase P activity found in the flow-through of anti-Sm affinity columns can be separated into inactive RNA and protein components. These components can be used to reconstitute active hybrid enzyme complexes with purified subunits from E. coli RNAase P. The RNA in the HeLa cell fractions employed is enriched for species between 85 and 115 nucleotides long. This reconstitution assay is a convenient means of purifying the functional RNA and protein of HeLa cell RNAase P. Probes derived from the genes for the subunits of E. coli RNAase P hybridize to genomic DNA of gram-negative prokaryotic organisms, but no positive signals are seen with genomic DNA from a variety of eukaryotic organisms.",

author = "Gold, {Heidi A.} and Sidney Altman",

note = "Funding Information: We are grateful to Cecilia Guerrier-Takada, Charles Hart, and Agustin Vioque for many valuable discussions and gifts of material. Members of Joan Steitz{\textquoteright}s Laboratory freely contributed material and advice for the use of monoclonal antibodies. We also thank Nathan P Lawrence and Madeline Baer for their advice and Donna Wesolowski for her technical assistance. Our colleagues at Yale generously donated genomic DNAs. This work was supported by a National Institutes of Health predoctoral training fellowship to H. A. G. and by a grant from the National Science Foundation to S. A.",

year = "1986",

month = jan,

day = "31",

doi = "10.1016/0092-8674(86)90758-0",

language = "English (US)",

volume = "44",

pages = "243--249",

journal = "Cell",

issn = "0092-8674",

publisher = "Cell Press",

number = "2",

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TY - JOUR

T1 - Reconstitution of RNAase P activity using inactive subunits from E. coli and HeLa cells

AU - Gold, Heidi A.

AU - Altman, Sidney

N1 - Funding Information: We are grateful to Cecilia Guerrier-Takada, Charles Hart, and Agustin Vioque for many valuable discussions and gifts of material. Members of Joan Steitz’s Laboratory freely contributed material and advice for the use of monoclonal antibodies. We also thank Nathan P Lawrence and Madeline Baer for their advice and Donna Wesolowski for her technical assistance. Our colleagues at Yale generously donated genomic DNAs. This work was supported by a National Institutes of Health predoctoral training fellowship to H. A. G. and by a grant from the National Science Foundation to S. A.

PY - 1986/1/31

Y1 - 1986/1/31

N2 - HeLa cell RNAase P activity found in the flow-through of anti-Sm affinity columns can be separated into inactive RNA and protein components. These components can be used to reconstitute active hybrid enzyme complexes with purified subunits from E. coli RNAase P. The RNA in the HeLa cell fractions employed is enriched for species between 85 and 115 nucleotides long. This reconstitution assay is a convenient means of purifying the functional RNA and protein of HeLa cell RNAase P. Probes derived from the genes for the subunits of E. coli RNAase P hybridize to genomic DNA of gram-negative prokaryotic organisms, but no positive signals are seen with genomic DNA from a variety of eukaryotic organisms.

AB - HeLa cell RNAase P activity found in the flow-through of anti-Sm affinity columns can be separated into inactive RNA and protein components. These components can be used to reconstitute active hybrid enzyme complexes with purified subunits from E. coli RNAase P. The RNA in the HeLa cell fractions employed is enriched for species between 85 and 115 nucleotides long. This reconstitution assay is a convenient means of purifying the functional RNA and protein of HeLa cell RNAase P. Probes derived from the genes for the subunits of E. coli RNAase P hybridize to genomic DNA of gram-negative prokaryotic organisms, but no positive signals are seen with genomic DNA from a variety of eukaryotic organisms.

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U2 - 10.1016/0092-8674(86)90758-0

DO - 10.1016/0092-8674(86)90758-0

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SN - 0092-8674

VL - 44

SP - 243

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JO - Cell

JF - Cell

IS - 2

ER -

Reconstitution of RNAase P activity using inactive subunits from E. coli and HeLa cells

Abstract

ASJC Scopus subject areas

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