Eukaryotic RNA polymerases: comparative subunit structures, immunological properties, and .alpha.-amanitin sensitivities of the class II enzymes from higher plants

Jendrisak, Jerry; Guilfoyle, Tom J.

doi:10.1021/bi00600a029

Cited by 72 publications

(30 citation statements)

References 22 publications

(25 reference statements)

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“…Gels were stained with Coomassie brilliant blue R and then destained by the method of Cleveland et al (18). Electrophoretic patterns of the maize chloroplast and type II nuclear enzymes were identical to those presented elsewhere (10)(11)(12)(13)(14)(15).…”

supporting

confidence: 76%

Peptide maps comparing subunits of maize chloroplast and type II nuclear DNA-dependent RNA polymerases.

Kidd¹,

Bogorad²

1979

Proc. Natl. Acad. Sci. U.S.A.

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Both one-and two-dimensional pe tide mapping techniques have been used to compare the 180-kilodalton subunits from maize chloroplast and type II nuclear DNAdependent RNA polymerases (nucleosidetriphosphate:RNA nucleotidyltransferase, EC 2.7.7.6). Despite their similar molecular mass, these 180-kilodalton subunits are not the same. Also, two-dimensional tryptic maps of the 160-, 43-, and 28-kilodalton polypeptides from maize type II nuclear RNA polymerase and of the 140-, 42-, and 27-kilodalton polypeptides from maize chloroplast RNA polymerase show that each of these six polypeptides is unique. Cells of eukaryotic green plants contain at least three separate genomes. DNA is found together with DNA-dependent RNA polymerases (nucleosidetriphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) in their chloroplasts and mitochondria as well as in their nuclei (1-4). The maintenance and development of each organelle, as well as of the cell as a whole, require the participation of all three genomes. For example, ribosomes and other multimeric components of organelles are composed of organelle-and nuclear-coded elements (4). So, not only does RNA synthesis have to be regulated within each organelle, but also gene expression in such multigenomic cells must be coordinated and integrated. Intracellular integration of transcription could be effected through functional ties between nuclear and organellar RNA polymerases. Chloroplast and type II nuclear RNA polymerases have been prepared from subcellular fractions of maize leaf homogenates (5-14). Sodium dodecyl sulfate/polyacrylamide gel electrophoresis of these two maize RNA polymerases has shown that each enzyme contains two large subunits which are present at all stages of purification. The molecular masses of these polypeptides are 180 and 160 kilodaltons (kDal) for the type II nuclear RNA polymerase (12, 14, 15) and 180 and 140 kDal for the chloroplast enzyme (11). Moreover, when a sample containing both the plastid and nuclear enzymes is electrophoresed through a single denaturing gel, the 180-kDal subunits of the two RNA polymerases are not resolved (12). Intracellular-intergenomic integration of transcription via sharing of a common subunit by chloroplast and type II nuclear RNA polymerases seems an appealing possibility.In this paper we compare, by one-and two-dimensional mapping, proteolytic fragments of the 180-kDal subunits as well as other pairs of polypeptides from maize chloroplast and type II nuclear RNA polymerases. None of the pairs of subunits of the two enzymes resembles one another. MATERIALS AND METHODSPlants. Maize seeds (Zea mays, FR9c ms X FR37, Illinois Foundation Seeds) were soaked in water, then sown in moist vermiculite, and grown in a greenhouse for 7-9 days with supplemental light.RNA Polymerases and Subunits. Chloroplast RNA polymerase was solubilized from isolated maize chloroplasts (7) and was purified by glycerol gradient centrifugation and phosphocellulose chromatography (11, 13). Type II nuclear RNA polymerase was purified as describe...

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supporting

confidence: 76%

Peptide maps comparing subunits of maize chloroplast and type II nuclear DNA-dependent RNA polymerases.

Kidd¹,

Bogorad²

1979

Proc. Natl. Acad. Sci. U.S.A.

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“…4A). The subunit composition of Arabidopsis RNA polymerase II is highly similar to other plant RNA polymerase II enzymes, with each enzyme containing 11 subunits resolved in 15% SDS gels (39). To obtain better resolution of the three largest subunits (i.e.…”

Section: Resultsmentioning

confidence: 99%

“…The B36b subunit may be more abundant in Arabidopsis RNA polymerase II found in specific tissues or specific developmental stages. It is worth noting that heterogeneity in the size of the third largest subunit in RNA polymerase II has been reported for enzymes purified from wheat and rye embryos (39,42), suggesting that more than one gene encodes this subunit in other plant species.…”

Section: Discussionmentioning

confidence: 99%

Association between 36- and 13.6-kDa α-Like Subunits of Arabidopsis thaliana RNA Polymerase II

Ulmasov

Larkin

Guilfoyle

1996

Journal of Biological Chemistry

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Two subunits in RNA polymerase II (e.g. RPB3 and RPB11 in yeast) and two subunits common to RNA polymerases I and III (e.g. AC40 and AC19 in yeast) contain one or two motifs related to the ␣ subunit in prokaryotic RNA polymerases. We have sequenced two different cDNAs (AtRPB36a and AtRPB36b), the two corresponding genes from Arabidopsis thaliana that are homologs of yeast RPB3, and an Arabidopsis cDNA (AtRPB13.6) that is a homolog of yeast RPB11. The B36a subunit is the predominant B36 subunit associated with RNA polymerase II purified from Arabidopsis suspension culture cells, and this subunit has a stoichiometry of about 1. Results from protein association assays showed that the B36a and B36b subunits did not associate, but each of these subunits did associate with the B13.6 subunit in vivo and in vitro. Two motifs in the B36b subunit related to the prokaryotic ␣ subunit were shown to be required for the in vitro interactions with the B13.6 subunit. Our results suggest that the B36 and B13.6 subunits associate to form heterodimers in Arabidopsis RNA polymerase II like the AC40 and AC19 heterodimers reported for yeast RNA polymerases I and III but unlike the B44 homodimers reported for yeast RNA polymerase II.Eukaryotes contain three classes of nuclear RNA polymerase, referred to as RNA polymerases I or A, II or B, and III or C. Each class of RNA polymerase is a multimeric enzyme composed of two unique large subunits in excess of 100 kDa that are related to ␤Ј and ␤ subunits of Escherichia coli RNA polymerase and 10 or more smaller subunits (reviewed in Refs. 1-5). In the yeast Saccharomyces cerevisiae five of these smaller subunits are common to RNA polymerase I, II, and III, and seven subunits are common to RNA polymerases I and III (3, 4). Subunits of about 40 kDa (e.g. yeast AC40 and B44 or RPB3) and 12.5-19 kDa (e.g. yeast AC19 and B12.5 or RPB11) in RNA polymerase I, II, and III have limited amino acid sequence homology with the ␣ subunit of the prokaryotic RNA polymerase (6 -8). The localized amino acid sequence homology between the eukaryotic ␣-like subunits and the ␣ subunit in prokaryotic RNA polymerases has been referred to as the "␣ motif" (2, 9). The yeast AC40 and AC19 subunits are common to RNA polymerase I and III, and the related subunits, B44 or RPB3 and B12.5 or RPB11, are unique to RNA polymerase II (6,8,10,11). Bacterial RNA polymerase has an ␣ subunit with a stoichiometry of 2, and the core enzyme is composed of ␣ 2 ␤␤Ј (12). The yeast B44 subunit is reported to have a stoichiometry of 2 in RNA polymerase II (13), but the AC40 and AC19 subunits in RNA polymerases I and III have apparent stoichiometries of 1 (3, 14). The stoichiometry of the yeast B12.5 subunit has not been reported.Yeast RNA polymerase II contains a total of 12 subunits, and each of these is encoded by a single copy gene (reviewed in Refs. 3 and 4). All of the RNA polymerase II subunit genes in yeast have been sequenced. Only a limited number of RNA polymerase II subunit genes in other eukaryotes have been cloned and seque...

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“…Little noticeable difference in inhibition was observed with these three higher levels of alpha-amanitin; an average of 51% of the transcription was resistant. At these concentrations of alpha-amanitin to which RNA polymerase I is resistant (13), the total level of rRNA transcription in soybean nuclei was nearly identical in light and darkness.…”

Section: Methodsmentioning

confidence: 92%

Transcriptional regulation of a gene encoding the small subunit of ribulose-1,5-bisphosphate carboxylase in soybean tissue is linked to the phytochrome response.

Berry-Lowe¹,

Meagher²

1985

Mol. Cell. Biol.

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The effects of white light, far-red light, and darkness on the transcription of a soybean ribulose-1,5-biphosphate carboxylase smaUl subunit gene, SRS1, were investigated. RNA was labeled with [a-32PJUTP in nuclei isolated from plants grown under different conditions of light and darkness and used to probe Southern blots and dot blots. The levels of smaUl subunit mRNA synthesis were normalized to ribosomal RNA synthesis. We demonstrate that the SRS1 gene is transcribed at a rate 16-to 32-fold higher in plants grown in the light than in those grown in darkness. Transcription of the small subunit increased dramaticaly when plants grown in darkness were given 30 min to 6 h of light and then leveled off after 24 to 48 h of exposure. When light-grown seedlings were exposed to greater than 2 h of darkness, a gradual decrease in transcription was detected. This decrease in transcription reached basal dark-grown levels after 48 h of exposure to darkness. The increase in transcription in etiolated seedlings treated with white light for 15 min could be reduced to basal levels if the treatment was folowed by treatment with far-red light for 15 min. In addition, transcription in light-grown seedlings was reduced to basal levels when plants were exposed to far-red light for 15 min. The transcription of this ribulose-1,5-biphosphate carboxylase smal subunit gene is strongly positively regulated by white light, is negatively regulated by far-red light, and exhibits a classic phytochrome-linked response.

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Eukaryotic RNA polymerases: comparative subunit structures, immunological properties, and .alpha.-amanitin sensitivities of the class II enzymes from higher plants

Cited by 72 publications

References 22 publications

Peptide maps comparing subunits of maize chloroplast and type II nuclear DNA-dependent RNA polymerases.

Peptide maps comparing subunits of maize chloroplast and type II nuclear DNA-dependent RNA polymerases.

Association between 36- and 13.6-kDa α-Like Subunits of Arabidopsis thaliana RNA Polymerase II

Transcriptional regulation of a gene encoding the small subunit of ribulose-1,5-bisphosphate carboxylase in soybean tissue is linked to the phytochrome response.

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