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W Bottomley

Publications and source records attributed to W Bottomley.

26 records · Page 2Linked to original sources
PubMed1978

Low-molecular-weight (4.5S) ribonucleic acid in higher-plant chloroplast ribosomes.

A species of RNA that migrates on 10% (w/v) polyacrylamide gels between 5S and 4S RNA was detected in spinach chloroplasts. This RNA (referred to as 4.5 S RNA) was present in amounts equimolar to the 5S RNA and its molecular weight was estimated to be approx. 33 000. Fractionation of the chloroplast components showed that the 4.5S RNA was associated with the 50 S ribosomal subunit and that it could be removed by washing the ribosomes with a buffer containing 0.01 M-EDTA and 0.5 M-KCl. It did not appear to be a cleavage product of the labile 23 S RNA of spinach chloroplast ribosomes. When 125I-labelled 4.5 S RNA was hybridized to fragments of spinach chloroplast DNA produced by SmaI restriction endonuclease, a single fragment (mol.wt. 1.15 times 10(6)) became labelled. The same DNA fragment also hybridized to chloroplast 5 S RNA and part of the 23 S RNA. It was concluded that the coding sequence for 4.5 S RNA was part of, or immediately adjacent to, the rRNA-gene region in chloroplast DNA . A comparable RNA species was observed in chloroplasts of tobacco and pea leaves.

Chloroplasts↗
PubMed1978

Mapping of the ribosomal RNA genes on spinach chloroplast DNA.

Spinach chloroplast ribosomal RNAs have been hybridized to restriction endonuclease fragments of spinach chloroplast DNA. All three RNA species (23S, 16S and 5S) hybridized to a single large fragment when the DNA was digested with either Sall or Pstl. Hybridization of 23S RNA to fragments produced by Smal yielded two radioactive bands which corresponded to the bi-molar 2.5 X 10(6) and 1.15 X 10(6) Mr fragments. 16S RNA also hybridized to two, bi-molar Smal fragments (3.4 X 10(6) and 2.5 X 10(6) Mr) and 5S RNA hybridized to the 1.15 X 10(6) Mr bi-molar Smal fragment. The 23S RNA and 16S RNA cistrons were each also shown to contain a single EcoRI site. From the data it was possible to conclude that the ribosomal RNA genes are located on the inverted repeat region of the spinach chloroplast DNA restriction map [1,2], that the sequence of the cistrons is 16S - 23S - 5S and that the size of the spacer between the 16S and 23S RNA cistrons is approximately 0.90 X 10(6) Mr.

Chloroplasts↗
PubMed1971

RNA polymerases of maize: partial purification and properties of the chloroplast enzyme.

A DNA-dependent RNA polymerase has been solubilized and partially purified from washed chloroplasts prepared from maize seedlings. The purified enzyme was completely dependent on added DNA after purification by phospho- or DEAE-cellulose chromatography. On glycerol density gradients, the enzyme ran ahead of a marker with a sedimentation constant of 18 S, indicating a molecular weight of 500,000 or more. The instability of the highly purified enzyme made intensive study difficult, but the properties of the enzyme purified by phosphocellulose chromatography are reported. Template specificity varied during purification but the activity was always higher with denatured than with native DNA and the preference for maize DNA over calf-thymus DNA increased during purification. The enzyme required magnesium for optimal activity, was inhibited by salt concentrations in excess of 0.1 M, and had a temperature optimum of 48 degrees C. The chloroplast enzyme differed from similar activities so far reported from maize or other sources, particularly in the high salt concentrations needed to elute it from phosphocellulose. The soluble, DNA-dependent enzyme was not inhibited by either alpha-amanitin or by rifamycin-SV under the assay conditions used.

Adenosine Monophosphate↗