Functional analysis of two maize cDNAs encoding T7-like RNA polymerases

Ching Chun Chang, Jen Sheen, Muriel Bligny, Yasuo Niwa, Silva Lerbs-Mache, David B. Stern

Research output: Contribution to journalArticle

95 Citations (Scopus)

Abstract

We have characterized two maize cDNAs, rpoTm and rpoTp, that encode putative T7-like RNA polymerases. In vivo cellular localization experiments using transient expression of the green fluorescent protein suggest that their encoded proteins are targeted exclusively to mitochondria and plastids, respectively. An antibody raised against the C terminus of the rpoTp gene product identified mitochondrial polypeptides of ~100 kD. Their presence was correlated with RNA polymerase activity, and the antibody inhibited mitochondrial in vitro transcription activity. Together, these results strongly suggest that the product of rpoTm is involved in maize mitochondrial transcription. By contrast, immunoblot analysis and an antibody-linked polymerase assay indicated that rpoTp specifies a plastid RNA polymerase component. A quantitative reverse transcription-polymerase chain reaction assay was used to study the transcription of rpoTp and rpoTm in different tissues and under different environmental conditions. Although both genes were constitutively expressed, rpoTm transcripts were generally more prevalent in nonphotosynthetic tissues, whereas an increase in rpoTp transcripts paralleled chloroplast development. We suggest that these two genes encode constitutive components of the organelle transcription machinery but that their expression is nonetheless subject to modulation during plant development.

Original languageEnglish
Pages (from-to)911-926
Number of pages16
JournalPlant Cell
Volume11
Issue number5
DOIs
Publication statusPublished - 1999 May 1

Fingerprint

DNA-directed RNA polymerase
Zea mays
Plastids
Complementary DNA
transcription (genetics)
DNA-Directed RNA Polymerases
corn
Antibodies
Genes
antibodies
plastids
Plant Development
Chloroplasts
Green Fluorescent Proteins
Organelles
Reverse Transcription
genes
Mitochondria
assays
green fluorescent protein

All Science Journal Classification (ASJC) codes

  • Plant Science
  • Cell Biology

Cite this

Chang, C. C., Sheen, J., Bligny, M., Niwa, Y., Lerbs-Mache, S., & Stern, D. B. (1999). Functional analysis of two maize cDNAs encoding T7-like RNA polymerases. Plant Cell, 11(5), 911-926. https://doi.org/10.1105/tpc.11.5.911
Chang, Ching Chun ; Sheen, Jen ; Bligny, Muriel ; Niwa, Yasuo ; Lerbs-Mache, Silva ; Stern, David B. / Functional analysis of two maize cDNAs encoding T7-like RNA polymerases. In: Plant Cell. 1999 ; Vol. 11, No. 5. pp. 911-926.
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Chang, CC, Sheen, J, Bligny, M, Niwa, Y, Lerbs-Mache, S & Stern, DB 1999, 'Functional analysis of two maize cDNAs encoding T7-like RNA polymerases', Plant Cell, vol. 11, no. 5, pp. 911-926. https://doi.org/10.1105/tpc.11.5.911

Functional analysis of two maize cDNAs encoding T7-like RNA polymerases. / Chang, Ching Chun; Sheen, Jen; Bligny, Muriel; Niwa, Yasuo; Lerbs-Mache, Silva; Stern, David B.

In: Plant Cell, Vol. 11, No. 5, 01.05.1999, p. 911-926.

Research output: Contribution to journalArticle

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Chang CC, Sheen J, Bligny M, Niwa Y, Lerbs-Mache S, Stern DB. Functional analysis of two maize cDNAs encoding T7-like RNA polymerases. Plant Cell. 1999 May 1;11(5):911-926. https://doi.org/10.1105/tpc.11.5.911