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Long distance movement of an Arabidopsis Translationally Controlled Tumor Protein (AtTCTP2) mRNA and protein in tobacco.

Toscano-Morales R, Xoconostle-Cázares B, Martínez-Navarro AC, Ruiz-Medrano R - Front Plant Sci (2014)

Bottom Line: The results indicate that both AtTCTP2 mRNA and protein are capable of moving long distance in both directions (stock-scion and scion-stock) with a tendency for movement from source to sink tissue (stock to scion).In addition, the protein localization pattern in transgenic aerial and primary roots was basically the same, indicating specific nuclear destination in roots, but also in leaves.These findings provide an approach to understand the role of long-distance movement in the function of plant TCTPs, supporting the notion that some of these act in a non-cell autonomous manner, as the human counterpart, the Histamine Releasing Factor (HRF).

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology and Bioengineering, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional Mexico, Mexico.

ABSTRACT
Translationally Controlled Tumor Protein (TCTP) is an almost ubiquitous protein found in eukaryotes, fundamental for the regulation of development and general growth. The multiple functions of TCTP have been inferred from its involvement in several cell pathways, but the specific function of TCTP is still not known in detail. On the other hand, TCTP seems to respond to a plethora of external signals, and appears to be regulated at the transcriptional and/or translational levels by mechanisms yet to be determined. In the present work, we analyzed the capacity of AtTCTP2 gene products (mRNA and protein) to translocate long distance through tobacco heterografts (transgenic/WT and WT/transgenic). The results indicate that both AtTCTP2 mRNA and protein are capable of moving long distance in both directions (stock-scion and scion-stock) with a tendency for movement from source to sink tissue (stock to scion). Interestingly, aerial roots emerged only in heterografts where the protein was detected in both stock and scion, suggesting a correlation between the presence of AtTCTP2 and aerial root appearance. More detailed analysis showed that these aerial roots harbored the transgene and expressed both transcript and protein. In addition, the protein localization pattern in transgenic aerial and primary roots was basically the same, indicating specific nuclear destination in roots, but also in leaves. These findings provide an approach to understand the role of long-distance movement in the function of plant TCTPs, supporting the notion that some of these act in a non-cell autonomous manner, as the human counterpart, the Histamine Releasing Factor (HRF).

No MeSH data available.


Related in: MedlinePlus

Detection of AtTCTP2-GFP mRNA long-distance movement in tobacco grafts. Total RNA samples from stocks and scions (transgenic or WT) were extracted and used as templates to perform RT-PCR detection of the transcript AtTCTP2-GFP. For further propose we denote grafts as scion (apex) genotype/rootstock genotype. (A) In WT/AtTCTP2 grafts the mRNA of AtTCTP2-GFP was detected in half of the wt scions (see Table 1), while in (B) AtTCTP2/WT grafts the transcript was detected in one wt stock from the seven grafts performed (see Table 1). (C) Homografts WT/WT were carried out as controls, and no signal was detected. RT-PCR detection of 18S was performed as control for RNA quality in all samples (below A–C).
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Figure 1: Detection of AtTCTP2-GFP mRNA long-distance movement in tobacco grafts. Total RNA samples from stocks and scions (transgenic or WT) were extracted and used as templates to perform RT-PCR detection of the transcript AtTCTP2-GFP. For further propose we denote grafts as scion (apex) genotype/rootstock genotype. (A) In WT/AtTCTP2 grafts the mRNA of AtTCTP2-GFP was detected in half of the wt scions (see Table 1), while in (B) AtTCTP2/WT grafts the transcript was detected in one wt stock from the seven grafts performed (see Table 1). (C) Homografts WT/WT were carried out as controls, and no signal was detected. RT-PCR detection of 18S was performed as control for RNA quality in all samples (below A–C).

Mentions: Transgenic tobacco plants harboring the overexpression construct 35S:AtTCTP2-GFP-TNOS were obtained using the regeneration protocol described by Hinojosa-Moya et al. (2013). Taking advantage of this, the progeny of these regenerated plants was selected for herbicide resistance and tested for presence of trangene by PCR. Transgenic and WT plants of similar size and developmental stage were used to perform reciprocal heterografting experiments as well as WT homografts. Total RNA was extracted from young and mature leaves in all grafting experiments; these were used as templates to perform the RT-PCR detection of the AtTCTP2-GFP transcript to test for long-distance movement. Signal corresponding to GFP was detected in three out of six WT scions grafted onto transgenic stocks (Table 1), indicating long-distance movement of this mRNA (Figure 1A). Interestingly, in the case of the reciprocal heterograft, this signal was identified in one of seven grafts (Table 1, Figure 1B) suggesting directionality of the movement of this mRNA, likely from stock to scion (AtTCTP2-GFP/WT). The control homografts (WT/WT) did not show any transgenic signal (Figure 1C); 18S RNA was detected in all the samples at similar levels.


Long distance movement of an Arabidopsis Translationally Controlled Tumor Protein (AtTCTP2) mRNA and protein in tobacco.

Toscano-Morales R, Xoconostle-Cázares B, Martínez-Navarro AC, Ruiz-Medrano R - Front Plant Sci (2014)

Detection of AtTCTP2-GFP mRNA long-distance movement in tobacco grafts. Total RNA samples from stocks and scions (transgenic or WT) were extracted and used as templates to perform RT-PCR detection of the transcript AtTCTP2-GFP. For further propose we denote grafts as scion (apex) genotype/rootstock genotype. (A) In WT/AtTCTP2 grafts the mRNA of AtTCTP2-GFP was detected in half of the wt scions (see Table 1), while in (B) AtTCTP2/WT grafts the transcript was detected in one wt stock from the seven grafts performed (see Table 1). (C) Homografts WT/WT were carried out as controls, and no signal was detected. RT-PCR detection of 18S was performed as control for RNA quality in all samples (below A–C).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4269120&req=5

Figure 1: Detection of AtTCTP2-GFP mRNA long-distance movement in tobacco grafts. Total RNA samples from stocks and scions (transgenic or WT) were extracted and used as templates to perform RT-PCR detection of the transcript AtTCTP2-GFP. For further propose we denote grafts as scion (apex) genotype/rootstock genotype. (A) In WT/AtTCTP2 grafts the mRNA of AtTCTP2-GFP was detected in half of the wt scions (see Table 1), while in (B) AtTCTP2/WT grafts the transcript was detected in one wt stock from the seven grafts performed (see Table 1). (C) Homografts WT/WT were carried out as controls, and no signal was detected. RT-PCR detection of 18S was performed as control for RNA quality in all samples (below A–C).
Mentions: Transgenic tobacco plants harboring the overexpression construct 35S:AtTCTP2-GFP-TNOS were obtained using the regeneration protocol described by Hinojosa-Moya et al. (2013). Taking advantage of this, the progeny of these regenerated plants was selected for herbicide resistance and tested for presence of trangene by PCR. Transgenic and WT plants of similar size and developmental stage were used to perform reciprocal heterografting experiments as well as WT homografts. Total RNA was extracted from young and mature leaves in all grafting experiments; these were used as templates to perform the RT-PCR detection of the AtTCTP2-GFP transcript to test for long-distance movement. Signal corresponding to GFP was detected in three out of six WT scions grafted onto transgenic stocks (Table 1), indicating long-distance movement of this mRNA (Figure 1A). Interestingly, in the case of the reciprocal heterograft, this signal was identified in one of seven grafts (Table 1, Figure 1B) suggesting directionality of the movement of this mRNA, likely from stock to scion (AtTCTP2-GFP/WT). The control homografts (WT/WT) did not show any transgenic signal (Figure 1C); 18S RNA was detected in all the samples at similar levels.

Bottom Line: The results indicate that both AtTCTP2 mRNA and protein are capable of moving long distance in both directions (stock-scion and scion-stock) with a tendency for movement from source to sink tissue (stock to scion).In addition, the protein localization pattern in transgenic aerial and primary roots was basically the same, indicating specific nuclear destination in roots, but also in leaves.These findings provide an approach to understand the role of long-distance movement in the function of plant TCTPs, supporting the notion that some of these act in a non-cell autonomous manner, as the human counterpart, the Histamine Releasing Factor (HRF).

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology and Bioengineering, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional Mexico, Mexico.

ABSTRACT
Translationally Controlled Tumor Protein (TCTP) is an almost ubiquitous protein found in eukaryotes, fundamental for the regulation of development and general growth. The multiple functions of TCTP have been inferred from its involvement in several cell pathways, but the specific function of TCTP is still not known in detail. On the other hand, TCTP seems to respond to a plethora of external signals, and appears to be regulated at the transcriptional and/or translational levels by mechanisms yet to be determined. In the present work, we analyzed the capacity of AtTCTP2 gene products (mRNA and protein) to translocate long distance through tobacco heterografts (transgenic/WT and WT/transgenic). The results indicate that both AtTCTP2 mRNA and protein are capable of moving long distance in both directions (stock-scion and scion-stock) with a tendency for movement from source to sink tissue (stock to scion). Interestingly, aerial roots emerged only in heterografts where the protein was detected in both stock and scion, suggesting a correlation between the presence of AtTCTP2 and aerial root appearance. More detailed analysis showed that these aerial roots harbored the transgene and expressed both transcript and protein. In addition, the protein localization pattern in transgenic aerial and primary roots was basically the same, indicating specific nuclear destination in roots, but also in leaves. These findings provide an approach to understand the role of long-distance movement in the function of plant TCTPs, supporting the notion that some of these act in a non-cell autonomous manner, as the human counterpart, the Histamine Releasing Factor (HRF).

No MeSH data available.


Related in: MedlinePlus