Papers by Sandrine Bonhomme
Gene, 2013
Keywords: Arabidopsis thaliana UBX-containing protein p97/VCP/CDC48 Gametophyte p97/CDC48 is a ma... more Keywords: Arabidopsis thaliana UBX-containing protein p97/VCP/CDC48 Gametophyte p97/CDC48 is a major AAA-ATPase that acts in many cellular events such as ubiquitin-dependent degradation and membrane fusion. Its specificity depends on a set of adaptor proteins, most of them containing the ubiquitin regulatory X (UBX) domain. Using a differential hybridization system, we isolated a UBX-containing protein that is expressed during the early phase of male gametophyte development in the crop Brassica napus and isolated and characterized its closest Arabidopsis thaliana homolog, AtPUX7. The AtPUX7 gene is expressed broadly in both the sporophyte and gametophyte due to regulation inferred by its first intron. The subcellular localization of AtPUX7 was assigned mainly to the nucleus in both the sporophyte and in pollen, mirroring the AAA-ATPase AtCDC48A localization. Furthermore, AtPUX7 interacts specifically with AtCDC48A in yeast as well as in planta in the nucleus. This interaction was mediated through the AtPUX7 UBX domain, which is located at the protein C-terminus, while an N-terminal UBA domain mediated its interaction with ubiquitin. Consistent with those results, a yeast-three hybrid analysis showed that AtPUX7 can act as a bridge between AtCDC48A and ubiquitin, suggesting a role in targeted protein degradation. It is likely that AtPUX7 acts redundantly with other members of the Arabidopsis PUX family because a null Atpux7-1 mutant does not display obvious developmental defects.
Molecular and General Genetics, 1994
Transcription of a putative mitochondrial gene (orf138) has previously been correlated with Ogura... more Transcription of a putative mitochondrial gene (orf138) has previously been correlated with Ogura cytoplasmic male-sterility (CMS) in rapeseed cybrids. In this paper, studies performed on a Brassica cybrid with a different organization of the orf138 locus confirm this association. We also show that mitochondria isolated from male-sterile rapeseed plants synthesize a polypeptide of 19 kDa, which is absent in fertile revertants. Antibodies against a glutathione S-transferase-ORF138 fusion protein were raised to establish that this 19 kDa polypeptide is the product of orf138. The anti-ORF138 serum was used to demonstrate that the orf138 translation product occurs only in sterile cybrids and co-purifies with the mitochondrial membrane fraction.
Development, 2015
Strigolactones (SLs), first identified for their role in parasitic and symbiotic interactions in ... more Strigolactones (SLs), first identified for their role in parasitic and symbiotic interactions in the rhizosphere, constitute the most recently discovered group of plant hormones. They are best known for their role in shoot branching but, more recently, roles for SLs in other aspects of plant development have emerged. In the last five years, insights into the SL biosynthetic pathway have also been revealed and several key components of the SL signaling pathway have been identified. Here, and in the accompanying poster, we summarize our current understanding of the SL pathway and discuss how this pathway regulates plant development.
Current Genetics, 1991
Spontaneous reversion to fertility was studied in the progeny of a cytoplasmic male-sterile (CMS)... more Spontaneous reversion to fertility was studied in the progeny of a cytoplasmic male-sterile (CMS) Brassica napus cybrid containing recombinant B. napus/ Ogura radish mitochondrial genomes. This reversion is concomitant with the disappearance of a 2.5 kb NcoI fragment present in the mitochondrial DNA of Ogura radish, and of CMS cybrids derived from plants carrying Ogura cytoplasm, and absent in the mitochondrial genome of normal Brassicas and fertile cybrids. This specific fragment hybridizes to a 1.4 kb transcript found only in male-sterile plants bearing an Ogura derived cytoplasm.
Current opinion in plant biology
Strigolactones (SLs), a group of small carotenoid-derived molecules, were first known for their f... more Strigolactones (SLs), a group of small carotenoid-derived molecules, were first known for their function in the rhizosphere in both symbiotic and parasitic interactions. Most of the progress for deciphering SL biosynthesis and signalling pathways comes from the use of high branching mutants identified in several species demonstrating that SLs also play a hormonal role in plant development. How SLs are perceived by the different organisms on which they show bioactivity is a current major challenge for the growing SL research community. These molecules very likely predate the colonization of land by plants and represent a fascinating example of signalling molecules involved in key innovations during plant evolution.
Genetics, 2001
Mutant analysis represents one of the most reliable approaches to identifying genes involved in p... more Mutant analysis represents one of the most reliable approaches to identifying genes involved in plant development. The screening of the Versailles collection of Arabidopsis thaliana T-DNA insertion transformants has allowed us to isolate different mutations affecting male gametophytic functions and viability. Among several mutated lines, five have been extensively studied at the genetic, molecular, and cytological levels. For each mutant, several generations of selfing and outcrossing have been carried out, leading to the conclusion that all these mutations are tagged and affect only the male gametophyte. However, only one out of the five mutations is completely penetrant. A variable number of T-DNA copies has integrated in the mutant lines, although all segregate at one mutated locus. Two mutants could be defined as "early mutants": the mutated genes are presumably expressed during pollen grain maturation and their alteration leads to the production of nonfunctional polle...
Current topics in microbiology and immunology, 1995
PLoS ONE, 2014
In vascular plants, strigolactones (SLs) are known for their hormonal role and for their role as ... more In vascular plants, strigolactones (SLs) are known for their hormonal role and for their role as signal molecules in the rhizosphere. SLs are also produced by the moss Physcomitrella patens, in which they act as signaling factors for controlling filament extension and possibly interaction with neighboring individuals. To gain a better understanding of SL action at the cellular level, we investigated the effect of exogenously added molecules (SLs or analogs) in moss growth media. We used the previously characterized Ppccd8 mutant that is deficient in SL synthesis and showed that SLs affect moss protonema extension by reducing caulonema cell elongation and mainly cell division rate, both in light and dark conditions. Based on this effect, we set up bioassays to examine chemical structure requirements for SL activity in moss. The results suggest that compounds GR24, GR5, and 5-deoxystrigol are active in moss (as in pea), while other analogs that are highly active in the control of pea branching show little activity in moss. Interestingly, the karrikinolide KAR 1 , which shares molecular features with SLs, did not have any effect on filament growth, even though the moss genome contains several genes homologous to KAI2 (encoding the KAR 1 receptor) and no canonical homologue to D14 (encoding the SL receptor). Further studies should investigate whether SL signaling pathways have been conserved during land plant evolution.
Plant Reproduction, 2014
F-box protein genes family is one of the largest gene families in plants, with almost 700 predict... more F-box protein genes family is one of the largest gene families in plants, with almost 700 predicted genes in the model plant Arabidopsis. F-box proteins are key components of the ubiquitin proteasome system that allows targeted protein degradation. Transcriptome analyses indicate that half of these F-box protein genes are found expressed in microspore and/or pollen, i.e., during male gametogenesis.
Methods in Molecular Biology, 2012
In this chapter, we review the main organogenesis features and associated regulation processes of... more In this chapter, we review the main organogenesis features and associated regulation processes of the moss Physcomitrella patens (P. patens) , the model plant for the Bryophytes. We highlight how the study of this descendant of the earliest plant species that colonized earth, brings useful keys to understand the mechanisms that determine and control both vascular and non vascular plants organogenesis. Despite its simple morphogenesis pattern, P. patens still requires the fi ne tuning of organogenesis regulators, including hormone signalling, common to the whole plant kingdom, and which study is facilitated by a high number of molecular tools, among which the powerful possibility of gene targeting/replacement. The recent discovery of moss cells reprogramming capacity completes the picture of an excellent model for studying plant organogenesis.
Plant Cell Monographs, 2006
... Screening and Analysis of Pollen Tube Mutations Hélène Guermonprez · Fabien Nogué · Sandrine ... more ... Screening and Analysis of Pollen Tube Mutations Hélène Guermonprez · Fabien Nogué · Sandrine Bonhomme (u) ... traffick ing) yes L o bstein et al. 2004 hap3 PT early g ro w th SU C 1 (Su cro se tran sp o rter) no Jo hn so n et al. 2004 hap6 P T early g ro w th R ib o pho rinII n o ...
The Plant Journal, 2003
In higher plants, pollen tubes and root hairs share an ancient growth process named tip growth. W... more In higher plants, pollen tubes and root hairs share an ancient growth process named tip growth. We have isolated three allelic Arabidopsis mutant lines showing kinky-shaped pollen tubes and, when homozygous, showing shorter and thicker root hairs. The ultrastructure of pollen tubes in these kinky pollen (kip) mutants is similar to that of the wild type; however, time-lapse studies suggest that aberrant pollen tube shape is caused by periodic growth arrests alternated with phases of tube axis reorientation. The KIP gene encodes a protein of 2587 amino acids that is predicted to be targeted to the secretory pathway. KIP mRNA was detected in all organs investigated but was most abundant in pollen and roots. KIP has putative homologues in many eukaryotes, including mammals and yeast, and is similar to the Arabidopsis SABRE gene, whose mutation causes a dwarf phenotype. The phenotype of the kip/sab double mutant suggests related functions for both genes, however, the KIP protein is mostly required for tip-growth.
THE PLANT CELL ONLINE, 2009
We investigated the role of the ubiquitin proteasome system (UPS), which allows proteins to be se... more We investigated the role of the ubiquitin proteasome system (UPS), which allows proteins to be selectively degraded, during gametophyte development in Arabidopsis thaliana. Three mutant alleles altering the UPS were isolated in the Wassilewskija (Ws) accession: they affect the Regulatory Particle 5a (RPT5a) gene, which (along with RPT5b) encodes one of the six AAA-ATPases of the proteasome regulatory particle. In the heterozygous state, all three mutant alleles displayed 50% pollen lethality, suggesting that RPT5a is essential for male gametophyte development. However, a fourth mutant in the Columbia (Col) accession did not display such a phenotype because the RPT5b Col allele complements the rpt5a defect in the male gametophyte, whereas the RPT5b Ws allele does not. Double rpt5a rpt5b mutants showed a complete male and female gametophyte lethal phenotype in a Col background, indicating that RPT5 subunits are essential for both gametophytic phases. Mitotic divisions were affected in double mutant gametophytes correlating with an absence of the proteasome-dependent cyclinA3 degradation. Finally, we show that RPT5b expression is highly increased when proteasome functioning is defective, allowing complementation of the rpt5a mutation. In conclusion, RPT5 subunits are not only essential for both male and female gametophyte development but also display accession-dependent redundancy and are crucial in cell cycle progression.
PLANT PHYSIOLOGY, 2004
The screening of the Versailles collection of Arabidopsis T-DNA transformants allowed us to ident... more The screening of the Versailles collection of Arabidopsis T-DNA transformants allowed us to identify several male gametophytic mutants, including poky pollen tube (pok). The pok mutant, which could only be isolated as a hemizygous line, exhibits very short pollen tubes, explaining the male-specific transmission defect observed in this line. We show that the POK gene is duplicated in the Arabidopsis genome and that the predicted POK protein sequence is highly conserved from lower to higher eukaryotes. The putative POK homolog in yeast (Saccharomyces cerevisiae), referred to as Vps52p/SAC2, has been shown to be located at the late Golgi and to function in a complex with other proteins, Vps53p, Vps54p, and Vps51p. This complex is involved in retrograde trafficking of vesicles between the early endosomal compartment and the trans-Golgi network. We present the expression patterns of the POK gene and its duplicate P2 in Arabidopsis, and of the putative Arabidopsis homologs of VPS53 and VPS54 of yeast. We show that a POK::GFP fusion protein localizes to Golgi in plant cells, supporting the possibility of a conserved function for Vps52p and POK proteins. These results, together with the expression pattern of the POK::GUS fusion and the lack of plants homozygous for the pok mutation, suggest a more general role for POK in polar growth beyond the pollen tube elongation process. ; fax 33-1-30-83-33-19.
Plant Molecular Biology, 1989
The proton-translocating FIFo ATPase (ATP synthase) is one of the few enzyme complexes that has f... more The proton-translocating FIFo ATPase (ATP synthase) is one of the few enzyme complexes that has functionally equivalent mitochondrial and chloroplast counterparts encoded in the respective organellar DNAs. In particular, the plant mitochondrial (mt) genes, atpA, atp6, and atp9 are known to be homologous to the chloroplast (chl) atpA, atpI and atpH genes . Comparisons of the sequences of such genes from different plants should provide information about evolutionary processes within separate compartments of the same cell.
MGG Molecular & General Genetics, 1992
Sequence analysis of the Ogura-specific mitochondrial DNA (mtDNA) fragment isolated previously fr... more Sequence analysis of the Ogura-specific mitochondrial DNA (mtDNA) fragment isolated previously from Brassica cybrids carrying Ogura cytoplasmic male sterility (cms) revealed a tRNA fMot sequence, a putative 138 amino acid open reading frame (orf138), and a 158 amino acid ORF (orf158) previously observed in mitochondrial genomes from several other plant species. Transcription mapping showed that both ORFs are present on a 1.4 kb cms-specific transcript. The orf158 sequence is also transcribed in fertile plants on a different mRNA, and thus is unlikely to be related to cms. On the other hand, fertile revertant plants lack transcripts of the orf138 sequence, whose possible role in the mechanism of Ogura cms is discussed.
MGG Molecular & General Genetics, 1994
Transcription of a putative mitochondrial gene (orf138) has previously been correlated with Ogura... more Transcription of a putative mitochondrial gene (orf138) has previously been correlated with Ogura cytoplasmic male-sterility (CMS) in rapeseed cybrids. In this paper, studies performed on a Brassica cybrid with a different organization of the orf138 locus confirm this association. We also show that mitochondria isolated from male-sterile rapeseed plants synthesize a polypeptide of 19 kDa, which is absent in fertile revertants. Antibodies against a glutathione S-transferase-ORF138 fusion protein were raised to establish that this 19 kDa polypeptide is the product of orf138. The anti-ORF138 serum was used to demonstrate that the orf138 translation product occurs only in sterile cybrids and co-purifies with the mitochondrial membrane fraction.
Molecular and General Genetics MGG, 1998
Many genes are thought to be expressed during the haploid phase in plants, however, very few hapl... more Many genes are thought to be expressed during the haploid phase in plants, however, very few haploid-specific genes have been isolated so far. T-DNA insertion mutagenesis is a powerful tool for generating mutations that affect gametophyte viability and function, as disruption of a gene essential for these processes should lead to a defect in the transmission of the gametes. Mutants can therefore be screened on the basis of segregation distortion for a reporter resistance gene contained in the T-DNA. We have screened the Versailles collection of Arabidopsis transformants for 1:1 KanR:KanS segregation after selfing, focussing on gametophyte mutations which show normal transmission through one gametophyte and cause lethality or dysfunction of the other. Only 1.3% (207) of the 16,000 lines screened were scored as good candidates. Thorough genetic analysis of 38 putative T-DNA transmission defect lines (Ttd) identified 8 defective gametophyte mutants, which all showed 0 to 1% T-DNA transmission through the pollen. During the screen, we observed a high background of low-penetrance mutations, often affecting the function of both gametophytes, and many lines which were likely to carry chromosomal rearrangements. The reasons for the small number of retained lines (all male gametophytic) are discussed, as well as the finding that, for most of them, residual T-DNA transmission is obtained through the affected gametophyte.
Journal of Experimental Botany, 2008
The organization and dynamics of the plant endomembrane system require both universal and plant-s... more The organization and dynamics of the plant endomembrane system require both universal and plant-specific molecules and compartments. The latter, despite the growing wealth of information, remains poorly understood. From the study of an Arabidopsis thaliana male gametophytic mutant, it was possible to isolate a gene named POKY POLLEN TUBE (POK) essential for pollen tube tip growth. The similarity between the predicted POK protein sequence and yeast Vps52p, a subunit from the GARP/VFT complex which is involved in the docking of vesicles from the prevacuolar compartment to the Golgi apparatus, suggested that the POK protein plays a role in plant membrane trafficking. Genetic analysis of Arabidopsis mutants affecting AtVPS53 or AtVPS54 genes which encode putative POK partners shows a transmission defect through the male gametophyte for all lines, which is similar to the pok mutant. Using a combination of biochemical approaches and specific antiserum it has been demonstrated that the POK protein is present in phylogenetically divergent plant species, associated with membranes and belongs to a high molecular weight complex. Combination of immunolocalization studies and pharmacological approaches in different plant cells revealed that the POK protein associates with Golgi and post-Golgi compartments. The role of POK in post-Golgi endo-membrane trafficking and as a member of a putative plant GARP/VFT complex is discussed.
Gene, 2013
Keywords: Arabidopsis thaliana UBX-containing protein p97/VCP/CDC48 Gametophyte p97/CDC48 is a ma... more Keywords: Arabidopsis thaliana UBX-containing protein p97/VCP/CDC48 Gametophyte p97/CDC48 is a major AAA-ATPase that acts in many cellular events such as ubiquitin-dependent degradation and membrane fusion. Its specificity depends on a set of adaptor proteins, most of them containing the ubiquitin regulatory X (UBX) domain. Using a differential hybridization system, we isolated a UBX-containing protein that is expressed during the early phase of male gametophyte development in the crop Brassica napus and isolated and characterized its closest Arabidopsis thaliana homolog, AtPUX7. The AtPUX7 gene is expressed broadly in both the sporophyte and gametophyte due to regulation inferred by its first intron. The subcellular localization of AtPUX7 was assigned mainly to the nucleus in both the sporophyte and in pollen, mirroring the AAA-ATPase AtCDC48A localization. Furthermore, AtPUX7 interacts specifically with AtCDC48A in yeast as well as in planta in the nucleus. This interaction was mediated through the AtPUX7 UBX domain, which is located at the protein C-terminus, while an N-terminal UBA domain mediated its interaction with ubiquitin. Consistent with those results, a yeast-three hybrid analysis showed that AtPUX7 can act as a bridge between AtCDC48A and ubiquitin, suggesting a role in targeted protein degradation. It is likely that AtPUX7 acts redundantly with other members of the Arabidopsis PUX family because a null Atpux7-1 mutant does not display obvious developmental defects.
Uploads
Papers by Sandrine Bonhomme