A Rapidly Light‐induced Chloroplast Protein With a High Turnover Coded for by Pea Nuclear Dna

Abstract

Proteolytlc digestion of the 2 4 WO-M, precursor and the 11 OOO-Mr transported membrane protein Preparation nf gel slices containing the bands of interest Preparative gel electrophoreses of either supernatants from trlchloroac-tic acrd preclpltatron of translation products in vitro 1200 pl t r a n s l a t i o n mixi or chloroplast membranes after transpurt 1400 pl translation supernatant were transported) W e r e performed o n 10-15(. gradient polyacrylamide gels. The bands were localized by added or internal menbrane pratems and stainrng. I:,. addlflon, t'le radloactlvrty Of the selected slices was compared with their nerghbourrng bands; in the j a e e of the gel from transport, no contaminating radioactivity was present. according to Wieland and Geargopulos (291. After drying, the plates were treated w l t h enhance spray (New Engl.ind Nuclear) and exposed on Du Pont Cronex 2 f i l m . Proteolytlc digestion with Staphylococcus aureus protease YBGel slices of 1 cm lengtii 11/15 of a preparative gel) were prerncubated for 30 mrn at room temperature i n 0.1 M NH HC03 and 0.0016% SDS. After homogenization In 0.9 ml of the same but fresh medlum, the protelns were eluted by rncubatlon for 1 h aL 55OC. The extraction was repeated twice and the combined supernatants were lyaphrlired. S . aureus protease. ThP prntelni were dlgested and separated

The Regulation of Enzymes Involved in Chlorophyll Biosynthesis

“…These polypeptides have been termed early light-induced proteins (ELIPs ; . In etiolated barley and pea seedlings, their expression reaches a maximum in mRNA and protein concentrations after approximately 2-4 h illumination Meyer and Kloppstech, 1984;Grimm and Kloppstech, 1987). At that time, all of the Pchlide originally present in the photoactive PORA-Pchlide complex of the prolamellar bodies has become photoreduced to Chlide, the POR-degrading protease has reached its full activity and, thus, the PORA protein begins to break down rapidly.…”

Distribution of the early light‐inducible protein in the thylakoids of developing pea chloroplasts

“…One of these nuclear genes, most actively transcribed during the early phases of illumination, is the gene for the early light-inducible protein (ELIP). The occurrence of the protein has been shown in pea (Meyer and Kloppstech, 1984), barley , bean (Tavladoraki unpublished results) and tobacco plastids (Kristahl and Herzfeld, unpublished results) which indicates a general function in chloroplast development.…”

“…In pea, during the uptake process, the 24-kDa precursor is processed into a 17-kDa protein (Meyer and Kloppstech, 1984). The mature protein is localized in the thylakoid membrane.…”

Circadian Control of the Accumulation of mRNAs for Light- and Heat-Inducible Chloroplast Proteins in Pea (Pisum sativum L.)

“…21 During our studies on the control of chloroplast development a group of nuclear-coded, light-inducible chloroplast proteins have been detected which exhibit a number of striking properties (10,21). Their mRNAs rapidly accumulate upon illumination and were designated ELIPs.…”

“…Moreover, the maxima and the minima for expression of heat shock genes are different from those of the light-inducible proteins. MATERIALS (2,17,21) following the original protocols (24,28). Care was taken to translate limiting amounts of RNA to avoid competition between the mRNAs (18).…”