Plant physiological adaptations to the massive foreign protein synthesis occurring in recombinant chloroplasts.
Genetically engineered chloroplasts have an extraordinary capacity to accumulate recombinant proteins. We have investigated in tobacco (Nicotiana tabacum) the possible consequences of such additional products on several parameters of plant development and composition. Plastid transformants were analyzed that express abundantly either bacterial enzymes, alkaline phosphatase (PhoA-S and PhoA-L) and 4-hydroxyphenyl pyruvate dioxygenase (HPPD), or a green fluorescent protein (GFP). In leaves, the HPPD and GFP recombinant proteins are the major polypeptides and accumulate to higher levels than Rubisco. Nevertheless, these engineered metabolic sinks do not cause a measurable difference in growth rate or photosynthetic parameters. The total amino acid content of transgenic leaves is also not significantly affected, showing that plant cells have a limited protein biosynthetic capacity. Recombinant products are made at the expense of resident proteins. Rubisco, which constitutes the major leaf amino acid store, is the most clearly and strongly down-regulated plant protein. This reduction is even more dramatic under conditions of limited nitrogen supply, whereas recombinant proteins accumulate to even higher relative levels. These changes are regulated posttranscriptionally since transcript levels of resident plastid genes are not affected. Our results show that plants are able to produce massive amounts of recombinant proteins in chloroplasts without profound metabolic perturbation and that Rubisco, acting as a nitrogen buffer, is a key player in maintaining homeostasis and limiting pleiotropic effects.
Bally
Julia
J
Bayer CropScience, Bioscience, F-69263 Lyon cedex 09, France.
Nadai
Marie
M
Vitel
Maxime
M
Rolland
Anne
A
Dumain
Raphael
R
Dubald
Manuel
M
D000222:Adaptation, Physiological; D000596:Amino Acids / Q000378:metabolism; D002736:Chloroplasts / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism / Q000494:pharmacology; D018515:Plant Leaves / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000254:growth & development / Q000378:metabolism; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism*; D012273:Ribulose-Bisphosphate Carboxylase / Q000378:metabolism; D014026:Nicotiana / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
D000596:Amino Acids; D012333:RNA, Messenger; D011994:Recombinant Proteins; D012273:Ribulose-Bisphosphate Carboxylase; D009584:Nitrogen