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Different subcellular localization and trafficking properties of KNOX class 1 homeodomain proteins from rice

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Abstract

Genes of the KN1-like homeobox (KNOX) class 1 encode transcription factors involved in shoot apical meristem development and maintenance. We studied the subcellular localization of Green Fluorescent Protein-tagged rice KNOX proteins (Oskn1-3) after particle bombardment of onion and rice cells and after transformation of Arabidopsis and rice with constitutive and inducible expression constructs. In all test systems, the three rice KNOX proteins showed nuclear and cytoplasmic localization patterns. However, Oskn1 additionally showed in some cells a distribution over punctae moving randomly in the cytosol. Use of an inducible expression system indicated a nuclear presence of Oskn1 in cells of the shoot apical meristem and post-transcriptional down-regulation in early leaf primordia. Arabidopsis and rice test systems were used to study effects of plant hormones and auxin transport inhibition on KNOX protein localization. Application of GA3 or 1-NAA shifted protein localization completely to the cytoplasm and resulted in loss of the punctae formed by Oskn1. Conversely, NPA application induced a complete nuclear localization of the KNOX proteins. To study intercellular movement of the KNOX proteins we set up a novel co-bombardment assay in which trafficking of untagged KNOX proteins was visualized through the co-trafficking of green fluorescent or blue fluorescent marker proteins. In multiple independent experiments Oskn1 trafficked more extensively to neighboring cells than Oskn2 and Oskn3. Differences in the localization and trafficking properties of Oskn1, Oskn2 and Oskn3 correlate with differences in mRNA localization patterns and functional differences between the rice KNOX genes and their putative orthologues from other species.

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Abbreviations

1-NAA:

naphtalene-1-acetic acid

2,4-D:

2,4-dichlorophenoxy acidic acid

BAP:

benzylaminopurine

BFP:

blue fluorescent protein

DAPI:

4,6-diamidino-2-phenylindole

ER:

endoplasmic reticulum

GA:

gibberellic acid

GFP:

green fluorescent protein

IAA:

indole-3-acetic acid

KN1:

KNOTTED1

KNAT:

Arabidopsis thaliana

KNOX:

Knotted-1 like homeobox

NLS:

nuclear localization signal

NPA:

1-N-naphthylphthalamic acid

OSH:

Oryza sativa homeobox

OSKN:

Oryza sativa KNOX

SAM:

shoot apical meristem

SEL:

size exclusion limit

NCAPP1:

non-cell-autonomous pathway protein1

SHR:

SHORT-ROOT

STM:

SHOOTMERISTEMLESS

WUS:

WUSCHEL

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Author notes

  1. Suzanne J. H. Kuijt

    Present address: Max-Planck-Institut für Züchtungsforschung, Carl-von-Linneé Weg 10, 50829, Köln, Germany

  2. Enrico Scarpella

    Present address: Department of Biological Sciences, University of Alberta, CW405 Biological Sciences Building, T6G 2E9, Edmonton AB, Canada

Authors and Affiliations

  1. Institute of Biology, Leiden University, Wassenaarseweg 64, 2333, Leiden, AL, the Netherlands

    Suzanne J. H. Kuijt, Gerda E. M. Lamers, Saskia Rueb, Enrico Scarpella, Pieter B. F. Ouwerkerk, Herman P. Spaink & Annemarie H. Meijer

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  1. Suzanne J. H. Kuijt
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  2. Gerda E. M. Lamers
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Correspondence to Annemarie H. Meijer.

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Kuijt, S.J.H., Lamers, G.E.M., Rueb, S. et al. Different subcellular localization and trafficking properties of KNOX class 1 homeodomain proteins from rice. Plant Mol Biol 55, 781–796 (2004). https://doi.org/10.1007/s11103-005-1967-y

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