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Temperature conditional cAMP-requiring mutant strains ofChlamydomonas reinhardtii arrest in G1 and are rescued by added cAMP

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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.

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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

ɛ-cAMP:

1,N6-etheno-cAMP

EDTA:

ethylenediaminetetraacetic acid

EGTA:

ethylene glycol-bis(β-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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Author notes

  1. S. Sakuanrungsirikul

    Present address: Department of Agriculture, Horticultural Research Institute, Bangkok, Thailand

  2. J. D. I. Harper

    Present address: School of Biological Sciences, The University of Sydney, Sydney, New South Wales, Australia

Authors and Affiliations

  1. Plant Cell Biology Group, Research School of Biological Sciences, Australian National University, G.P.O. Box 475, 2601, Canberra, ACT, Australia

    S. Sakuanrungsirikul, C. H. Hocart, J. D. I. Harper, C. W. Parker & P. C. L. John

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  1. S. Sakuanrungsirikul
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  2. C. H. Hocart
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  3. J. D. I. Harper
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  4. C. W. Parker
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  5. P. C. L. John
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Sakuanrungsirikul, S., Hocart, C.H., Harper, J.D.I. et al. Temperature conditional cAMP-requiring mutant strains ofChlamydomonas reinhardtii arrest in G1 and are rescued by added cAMP. Protoplasma 192, 159–167 (1996). https://doi.org/10.1007/BF01273888

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  • DOI: https://doi.org/10.1007/BF01273888

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