Temperature conditional cAMP-requiring mutant strains ofChlamydomonas reinhardtii arrest in G1 and are rescued by added cAMP |
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Authors: | S Sakuanrungsirikul C H Hocart J D I Harper C W Parker P C L John |
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Institution: | (1) Plant Cell Biology Group, Research School of Biological Sciences, Australian National University, G.P.O. Box 475, 2601 Canberra, ACT, Australia;(2) Present address: Department of Agriculture, Horticultural Research Institute, Bangkok, Thailand;(3) Present address: School of Biological Sciences, The University of Sydney, Sydney, New South Wales, Australia |
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Abstract: | Summary Three independent isolates ofChlamydomonas, selected for caffeine resistance, were found to arrest in G1 phase, as determined by quantitative fluorescence measurements of DNA, when grown at a non-permissive temperature. This cell cycle arrest correlated with lowered levels of cAMP and of adenylate cyclase activity. The arrested cells could be rescued by added cAMP but not AMP, hence the defect was not one of general purine metabolism. Back-crosses to wild type revealed that the phenotypes observed result from a combination of three separable mutations. It is clear that the mutations define functions that are more stringently required for cell division than for growth since the mutant strains are able to grow up to fifteen times normal size while blocked at the non-permissive temperature. The possible interaction of cAMP dependent events with division is discussed.Abbreviations AMP
adenosine 5 -monophosphate
- ATP
adenosine 5 -triphosphate
- BSA
bovine serum albumin
- cAMP
adenosine 3 ,5 -cyclicmonophosphate
- db-cAMP
dibutyryl-cAMP
- DNA
deoxyribonucleic acid
- DTT
dithiothreitol
- -cAMP
1,N6-etheno-cAMP
- EDTA
ethylenediaminetetraacetic acid
- EGTA
ethylene glycol-bis( -aminoethylether)-N,N,N ,N -tetraacetic acid
- HPLC
high performance liquid chromatography
- LSA
low sulphur-high salt-acetate medium
- LYP LSA
media containing yeast extract and proteose peptone
- M1, 2, 3
mutants 1, 2, 3
- PDE
phosphodiesterase
- TAP
trisacetate-phosphate medium
- TLC
thin layer chromatography
- TYP TAP
medium containing yeast extract and proteose peptone |
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Keywords: | Adenylate cyclase Cell cycle Chlamydomonas reinhardtii Cyclic AMP Cyclic nucleotide phosphodiesterase Mutant strains |
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