In Vitro Protein Synthesis by Plastids of <i>Phaseolus vulgaris</i>. I. Localization of Activity in the Chloroplasts of a Chloroplast Containing Fraction from Developing Leaves

Parenti, Francesco; Margulies, Maurice M.

doi:10.1104/pp.42.9.1179

Cited by 44 publications

(32 citation statements)

References 24 publications

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“…Each ml of retaction mixtuire contained 0.8 ml of chloroplasts stuspended in Honda medium. In addition the following components were also present in 'the complete reaction mixtulre: ATP, pyruvate kinase, PEP, GTP, UTP, CTP, 12C-amino acids other than leucine, magnesium ion, ;and mercaptoethanol (13). Except where stated, incorporation was determined by scintillation counting by the method of Mans and Novdl'li (11).…”

Section: Methodsmentioning

confidence: 99%

“…A crude ch'loroplast preparation was utsed throutghouit (13 mince prepared by chopping leaves in Honda meditum (13,16). In addition to sucrose, tris, mercaptoethanol, and magnesitum ion, ithis meditum contains Ficoll and dextran.…”

Section: Methodsmentioning

confidence: 99%

“…Phaseoluts vulgaris L., var. Bla!ck Valentine was grown in the da,rk for 6.5 days, followed by 0.5 day of irradiation with white light (13).…”

Section: Methodsmentioning

confidence: 99%

“…When chloropliasts are suspended in, and in,corporation is carried otut in ihigh osmotic strength media, the reaction may be fully sensitive (16), partially sensitive (7,13), or insensitive (5) 'to ribontuclease. Insensitivity of a portion of inicorporaiting activity to high con'centrations of ribonuclease led to the suggestion that the insensitive amino acid incorporatilon was due to the presence of intact chloroplasts (13). The present work was carried ouit to investigate this possibility.…”

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confidence: 99%

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In vitro Protein Synthesis by Plastids of Phaseolus vulgaris. II. The Probable Relation Between Ribonuclease Insensitive Amino Acid Incorporation and the Presence of Intact Chloroplasts

1968

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Abstract. Amino acid incorporation into protein by chloroplasts from primary leaves of Phaseolus vulgaris L., var. Black Valentine is only partially inhibited by 400 ,uglml ribonuclease.The rate of incorporation, in the presence of ribonuclease, is progressively inhibited with time, and ceases after about half an hour. Preincubation of chloroplasts at 250, in the absence of ribonuclease, increases the inhibitory effect of ribonuclease on the initial rate of incorporation of amino acid into protein. Examination of electron micrographs of freshly prepared chloroplast suspensions shows that chloroplasts are largely intact. However, after incubation at 250 for 1 hour the chloroplasts are disrupted, as indicated by loss of their stroma oontents. It is concluded that the intact chloroplast membrane is relatively impermeable to ribonuclease. Amino acid incorporating activity probably becomes inhibited as the inside of the chloroplast is made accessible to ribonuclease by breakage of membranes during incubation at 25°.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Phaseoluts vulgaris L., var. Bla!ck Valentine was grown in the da,rk for 6.5 days, followed by 0.5 day of irradiation with white light (13).…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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In vitro Protein Synthesis by Plastids of Phaseolus vulgaris. II. The Probable Relation Between Ribonuclease Insensitive Amino Acid Incorporation and the Presence of Intact Chloroplasts

1968

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“…In addition, Triton X-100, which was used in the present experiments to aid in the liberation of ribosomes, is reported to solubilize plastids but not whole leaf cells, nuclei, or bacteria (1,14,30,34).…”

Section: Discussionmentioning

confidence: 99%

Effect of Light on Ribonucleic Acid Metabolism in Greening Maize Leaves

Harel¹,

Bogorad²

1973

Plant Physiol.

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The effect of illumination on the incorporation of labeled precursors into RNA of dark-grown maize (Zea niays) leaves was studied using either 3P-phosphate or double labeling with "4C-and 3H-uridine. In the dark, label was preferentially incorporated into etioplast ribosomal RNAs. Incorporation into this fraction and into lower molecular weight fractions was strongly and preferentially stimulated by light during the first 2 hours of illumination. The effect persisted after illumination was terminated. The possibility that light-induced alterations in plastid ribosomal RNA metabolism may not be required for chlorophyll accumulation in maize is discussed.Sucrose density gradient analyses of ribosomes and of extracted RNA did not reveal light-induced incorporation of label into messenger-like RNA associated with polyribosomes during brief illumination. However, newly produced RNA which seems to be neither ribosomal RNA nor transfer RNA is detectable after illumination for 2.5 hours or longer.Seedlings of angiosperms growing in complete darkness develop only immature chloroplasts which contain a small amount of protochlorophyllide but lack chlorophyll and many other components of the photosynthetic apparatus. Immediately upon illumination the protochlorophyllide is reduced to chlorophyllide, and the membranous elements of the prolamellar body are rearranged. Soon afterwards additional chlorophyll is produced and a number of plastid enzymes including RNA polymerase increase in activity. These changes are not all controlled by a single mechanism because the production of chlorophyll, for example, ceases soon after illuminated plants are returned to darkness but some other activities increase at accelerated rates even after illumination ceases. The role of light in regulating the various events which occur during greening is not understood although some segments of the processes have been described and studied in detail (3,5,6, 22).Investigations with inhibitors have suggested that continuous protein and DNA-dependent/RNA synthesis are required for at least some of the processes in the development of plastids in higher plants and in Euglena (2,7, 12. 16, 31. 33 MATERIALS AND METHODSMaize (Zea mays) seeds (WF 9 X B 37 -F1 single cross, Illinois Foundation Seeds, Inc.) were soaked for 1 day before planting and grown in vermiculite in the dark for 9 to 10 days at 27 C.Application of Isotopes, Preparation of Subceliular Fractions, and Isolation of Ribosomes. Leaves were harvested under a green safelight, and their cut bases were put into isotope solutions in small tubes. The leaves were placed in front of a fan to accelerate uptake of the isotope. Uptake in the dark usually required 1 to 1.5 hr. Plants were then either transferred to white fluorescent light (400-500 ft-c) or kept in the dark. At the end of treatment, the leaves were frozen in liquid N2 in the dark and were ground in a mortar and pestle with 0.5 ml of 0.5 M sucrose, 0.1 M tris-HCl buffer pH 7.6 containing 25 mM MgCl2 and 2 mm spermidine per gram ...

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The Synthesis of Chloroplast Membranes in Pisum sativum

Ellis

1975

Membrane Biogenesis

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In Vitro Protein Synthesis by Plastids of Phaseolus vulgaris. I. Localization of Activity in the Chloroplasts of a Chloroplast Containing Fraction from Developing Leaves

Cited by 44 publications

References 24 publications

In vitro Protein Synthesis by Plastids of Phaseolus vulgaris. II. The Probable Relation Between Ribonuclease Insensitive Amino Acid Incorporation and the Presence of Intact Chloroplasts

In vitro Protein Synthesis by Plastids of Phaseolus vulgaris. II. The Probable Relation Between Ribonuclease Insensitive Amino Acid Incorporation and the Presence of Intact Chloroplasts

Effect of Light on Ribonucleic Acid Metabolism in Greening Maize Leaves

The Synthesis of Chloroplast Membranes in Pisum sativum

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