Molecular biology of C4 photosynthesis in Zea mays: differential localization of proteins and mRNAs in the two leaf cell types

Richard Broglie, Gloria Coruzzi, Brian Keith, Nam Hai Chua

Research output: Contribution to journalArticle

Abstract

We have investigated the molecular basis of differential localization of enzyme activities in mesophyll(M) and bundle-sheath (B) cells of maize leaves. M protoplasts and B strands were prepared by enzymatic digestions and mechanical treatment of secondary leaves. Soluble and thylakoid membrane proteins from the two cell types were compared by one- and two-dimensional gel electrophoresis and quantitative rocket immunoelectrophoresis. In addition, several thylakoid polypeptides were identified by crossed immunoelectrophoresis using monospecific antibodies. M and B thylakoids show quantitative and qualitative differences in their polypeptide compositions. While the M thylakoids contain the normal complement of polypeptides, the B thylakoids are deficient in ferredoxin-NADP+ reductase, photosystem II reaction center polypeptides, and the light-harvesting chlorophyll a/b-protein complex. Comparison of the soluble proteins by two-dimensional gel electrophoresis revealed marked differences between M and B cells. The major proteins of one cell type are clearly absent from the other. These differences are paralleled by differences in the in vitro translation products of poly A+ RNA isolated from the two cell types. Immunoprecipitation experiments showed that mRNA encoding the small subunit of ribulose-1,5-bisphosphate carboxylase (rbcS) is localized exclusively in B cells, whereas mRNA encoding phosphoenolpyruvate carboxylase is detected only in M cells. cDNA clones encoding the carboxylase rbcS and the chlorophyll a/b binding protein were used as probes in Northern blot analysis. M cells contain no detectable RNA encoding rbcS but have a higher steady state level of RNA encoding the chlorophyll a/b-binding polypeptide compared to B cells. Taken together, our results demonstrate that differential gene expression in the two leaf cell types is regulated at the level of translatable mRNA, and, for at least two proteins, at the level of steady-state RNA.

Original languageEnglish (US)
Pages (from-to)431-444
Number of pages14
JournalPlant Molecular Biology
Volume3
Issue number6
DOIs
StatePublished - Nov 1984

Fingerprint

C4 photosynthesis
Molecular biology
Photosynthesis
molecular biology
Zea mays
Molecular Biology
thylakoids
polypeptides
Messenger RNA
Peptides
B-lymphocytes
Cells
chlorophyll
ribulose 1,5-diphosphate
B-Lymphocytes
RNA
Electrophoresis
Chlorophyll Binding Proteins
leaves
immunoelectrophoresis

ASJC Scopus subject areas

  • Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Molecular biology of C4 photosynthesis in Zea mays : differential localization of proteins and mRNAs in the two leaf cell types. / Broglie, Richard; Coruzzi, Gloria; Keith, Brian; Chua, Nam Hai.

In: Plant Molecular Biology, Vol. 3, No. 6, 11.1984, p. 431-444.

Research output: Contribution to journalArticle

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