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Disappearance of isocitrate lyase enzyme from cells of Chlorella pyrenoidosa.

ABSTRACT: 1. When acetate-adapted cells of Chlorella are suspended in nitrogen-free medium and supplied with glucose, isocitrate lyase activity disappears from the cells at a rate of about 9%/h. This loss of activity is shown to be accompanied by loss of isocitrate lyase protein. 2. When isocitrate lyase activity is assayed in intact cells after freezing and thawing, the rate of loss of activity after addition of glucose approaches 20%/h. 3. It is shown, by using (35)S, that the rate of turnover of isocitrate lyase protein is somewhat lower than that of other major soluble proteins; general protein turnover during nitrogen starvation, and after glucose addition, is too slow to account for the rate of loss of isocitrate lyase protein. 4. Disappearance of isocitrate lyase activity must result from a mechanism that allows degradation of this specific protein under conditions of limiting nitrogen supply.

SUBMITTER: John PC

PROVIDER: S-EPMC1179492 | biostudies-other | 1970 Oct

REPOSITORIES: biostudies-other

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Project description:BACKGROUND:Isocitrate lyase (ICL) is a key enzyme in the glyoxylate cycle. In a previous study in rice, the expression of the ICL-encoding gene (OsICL) was highly induced by salt stress and its expression was enhanced in transgenic rice lines overexpressing OsCam1-1, a calmodulin (CaM)-encoding gene. CaM has been implicated in salt tolerance mechanisms in plants; however, the cellular mechanisms mediated by CaM are not clearly understood. In this study, the role of OsICL in plant salt tolerance mechanisms and the possible involvement of CaM were investigated using transgenic plants expressing OsICL or OsCam1-1. RESULTS:OsICL was highly expressed in senesced leaf and significantly induced by salt stress in three OsCam1-1 overexpressing transgenic rice lines as well as in wild type (WT). In WT young leaf, although OsICL expression was not affected by salt stress, all three transgenic lines exhibited highly induced expression levels. In Arabidopsis, salt stress had negative effects on germination and seedling growth of the AtICL knockout mutant (Aticl mutant). To examine the roles of OsICL we generated the following transgenic Arabidopsis lines: the Aticl mutant expressing OsICL driven by the native AtICL promoter, the Aticl mutant overexpressing OsICL driven by the 35SCaMV promoter, and WT overexpressing OsICL driven by the 35SCaMV promoter. Under salt stress, the germination rate and seedling fresh and dry weights of the OsICL-expressing lines were higher than those of the Aticl mutant, and the two lines with the icl mutant background were similar to the WT. The Fv/Fm and temperature of rosette leaves in the OsICL-expressing lines were less affected by salt stress than they were in the Aticl mutant. Finally, glucose and fructose contents of the Aticl mutant under salt stress were highest, whereas those of OsICL-expressing lines were similar to or lower than those of the WT. CONCLUSIONS:OsICL, a salt-responsive gene, was characterized in the transgenic Arabidopsis lines, revealing that OsICL expression could revert the salt sensitivity phenotypes of the Aticl knockout mutant. This work provides novel evidence that supports the role of ICL in plant salt tolerance through the glyoxylate cycle and the possible involvement of OsCam1-1 in regulating its transcription.

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Disappearance of isocitrate lyase enzyme from cells of Chlorella pyrenoidosa.

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