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Phosphoenolpyruvate Carboxykinase in the C4 Monocot Urochloa panicoides Is Encoded by Four Differentially Expressed Genes1

Patrick M. Finnegan*, Shoichi Suzuki, Martha Ludwig, and James N. Burnell

Division of Biochemistry and Molecular Biology, Faculty of Science, Australian National University, Canberra, Australian Capital Territory 0200, Australia (P.M.F.); Japan Tobacco, 700 Higashibara Toyota-Cho Iwata Gun, Shizuoka 438-0802, Japan (S.S.); Department of Biological Sciences, Macquarie University, Sydney, New South Wales 2109, Australia (M.L.); and Department of Biochemistry and Molecular Biology, James Cook University, Townsville, Queensland 4810, Australia (J.N.B.)

Previous screening of a cDNA library of leaf poly(A+) RNA from Urochloa panicoides, a phosphoenolpyruvate carboxykinase (PCK)-type C4 monocot, led to the characterization of cDNAs encoding the U. panicoides PCK subunit PCK1. A second PCK sequence, designated PCK2, has now been found by rescreening the library. The deduced PCK2 polypeptide is 626 residues in length, has a predicted molecular mass of 68,686 D, and is 96% identical to the deduced PCK1 sequence. Isolation and characterization of genomic DNA fragments revealed that the PCK1 and PCK2 genes are each closely linked to another PCK gene. These additional genes have been designated PCK3 and PCK4, respectively. In each case, the second gene is located upstream and in the same transcriptional orientation as the gene characterized through cDNA analysis. A reverse transcription-polymerase chain reaction assay was used to demonstrate that PCK1 and PCK2 transcripts predominate in leaves, whereas PCK3 and PCK4 transcripts predominate in roots. Moreover, accumulation of PCK1 and PCK2 transcripts is light dependent. Direct N-terminal sequencing of PCK polypeptides purified from leaves demonstrated that PCK2 is produced. These results strongly suggest that PCK1 and PCK2 are involved in the photosynthetic CO2-concentrating mechanism active in U. panicoides.

1   This work was supported by a Joint Research and Development Grant from Japan Tobacco to J.N.B.
*   Corresponding author; e-mail Patrick.Finnegan{at}anu.edu.au; fax 61-2-6249-0313.

Plant Physiol. (1999) 120: 1033-1042
Copyright Clearance Center:   0032-0889/99/120//10
© 1999 American Society of Plant Physiologists




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