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Red Bell Pepper Chromoplasts Exhibit in Vitro Import Competency and Membrane Targeting of Passenger Proteins from the Thylakoidal Sec and Delta pH Pathways but Not the Chloroplast Signal Recognition Particle Pathway1

Elizabeth J. Summer and Kenneth Cline*

Horticultural Sciences Department and Plant Molecular and Cellular Biology Program, University of Florida, Gainesville, Florida 32611-0690

Chloroplast to chromoplast development involves new synthesis and plastid localization of nuclear-encoded proteins, as well as changes in the organization of internal plastid membrane compartments. We have demonstrated that isolated red bell pepper (Capsicum annuum) chromoplasts contain the 75-kD component of the chloroplast outer envelope translocon (Toc75) and are capable of importing chloroplast precursors in an ATP-dependent fashion, indicating a functional general import apparatus. The isolated chromoplasts were able to further localize the 33- and 17-kD subunits of the photosystem II O2-evolution complex (OE33 and OE17, respectively), lumen-targeted precursors that utilize the thylakoidal Sec and Delta pH pathways, respectively, to the lumen of an internal membrane compartment. Chromoplasts contained the thylakoid Sec component protein, cpSecA, at levels comparable to chloroplasts. Routing of OE17 to the lumen was abolished by ionophores, suggesting that routing is dependent on a transmembrane Delta pH. The chloroplast signal recognition particle pathway precursor major photosystem II light-harvesting chlorophyll a/b protein failed to associate with chromoplast membranes and instead accumulated in the stroma following import. The Pftf (plastid fusion/translocation factor), a chromoplast protein, integrated into the internal membranes of chromoplasts during in vitro assays, and immunoblot analysis indicated that endogenous plastid fusion/translocation factor was also an integral membrane protein of chromoplasts. These data demonstrate that the internal membranes of chromoplasts are functional with respect to protein translocation on the thylakoid Sec and Delta pH pathways.

1   This work was supported in part by National Institutes of Health grant no. R01 GM4691 (to K.C.). DNA sequencing was conducted by the University of Florida Interdisciplinary Center for Biotechnology Research (ICBR) DNA Sequencing Core, and electron microscopy was conducted by the ICBR Electron Microscopy Core Laboratory, both of which are supported by funds supplied by the Division of Sponsored Research and the ICBR at the University of Florida. This paper is Florida Agricultural Experiment Station journal series no. R-06592.
*   Corresponding author; e-mail KCC{at}nervm.nerdc.ufl.edu; fax 1-352-392-6479.

Plant Physiol. (1999) 119: 575-584
Copyright Clearance Center:   0032-0889/99/119//10
© 1999 American Society of Plant Physiologists




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