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Plant Cell, Vol. 13, 193-205, January 2001, Copyright © 2001, American Society of Plant Physiologists
The Gene for the Ribulose-1,5-Bisphosphate
Carboxylase/Oxygenase (Rubisco) Small Subunit Relocated to the Plastid Genome of Tobacco Directs the
Synthesis of Small Subunits That Assemble into Rubisco
Spencer M. Whitneya and
T. John Andrewsa
a Molecular Plant Physiology, Research School of Biological Sciences, Australian
National University, P.O. Box 475, Canberra 2601, Australia
Correspondence to:
T. John Andrews, john.andrews{at}anu.edu.au (E-mail), 61-2-6125-5075 (fax)
To assess the extent to which a
nuclear gene for a chloroplast protein retained the ability to be expressed in its presumed
preendosymbiotic location, we relocated the RbcS gene for the small subunit of
ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) to the tobacco plastid genome. Plastid
RbcS transgenes, both with and without the transit presequence, were equipped with 3'
hepta-histidine–encoding sequences and psbA promoter and terminator elements. Both
transgenes were transcribed abundantly, and their products were translated into small subunit
polypeptides that folded correctly and assembled into the Rubisco hexadecamer. When present, either
the transit presequence was not translated or the transit peptide was cleaved completely. After
assembly into Rubisco, transplastomic small subunits were relatively stable. The hepta-histidine
sequence fused to the C terminus of a single small subunit was sufficient for isolation of the whole
Rubisco hexadecamer by Ni2+ chelation. Small subunits produced by the plastid transgenes
were not abundant, never exceeding  1% of the total small subunits, and they differed from
cytoplasmically synthesized small subunits in their N-terminal modifications. The scarcity of
transplastomic small subunits might be caused by inefficient translation or
assembly.
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