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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

Comparison of the protein localization pattern between adventitious and primary roots in heterografts. Mid-root region from aerial or primary roots were used to identify AtTCTP2-GFP signal through fluorescence confocal microscopy. AtTCTP2-GFP nuclear localization (N) in the mid-root region of (A,D) aerial and (B,E) transgenic primary roots was recurrent in all cases, while in the (C,F) WT primary control no nuclear signal was found (N; dashed white circles). In addition, AtTCTP2-GFP was located in lateral root primordia (LRp) both in (G) aerial roots and (H) transgenic primary roots in contrast to (I) WT primary roots. Size bars: 50 μm.
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Figure 6: Comparison of the protein localization pattern between adventitious and primary roots in heterografts. Mid-root region from aerial or primary roots were used to identify AtTCTP2-GFP signal through fluorescence confocal microscopy. AtTCTP2-GFP nuclear localization (N) in the mid-root region of (A,D) aerial and (B,E) transgenic primary roots was recurrent in all cases, while in the (C,F) WT primary control no nuclear signal was found (N; dashed white circles). In addition, AtTCTP2-GFP was located in lateral root primordia (LRp) both in (G) aerial roots and (H) transgenic primary roots in contrast to (I) WT primary roots. Size bars: 50 μm.

Mentions: In addition, confocal fluorescence laser microscopy was performed to detect AtTCTP2-GFP protein signal and determine its localization as well. The AtTCTP2-GFP fluorescent signal was specifically found in the nuclei of the mid-root region in all aerial roots that emerged (Figure 6A) which is consistent with the localization pattern found in the 35S::AtTCTP2-GFP transgenic primary roots (Figure 6B; Toscano-Morales et al., submitted) and in contrast with the absence of nuclear signal in the primary mid-root region of WT stock controls (Figure 6C). Interestingly, no nuclear localization signal is predicted for AtTCTP2, or several plant TCTPs tested, using a nuclear predictor server (http://www.sbc.su.se/~maccallr/nucpred/). In a similar manner, the same localization pattern was observed in lateral root primordia both in aerial and transgenic primary root (Figures 6D,E,G,H), unlike the WT stock control (Figures 6F,I). Remarkably, the protein accumulation pattern (in cells at the base of the lateral root primordia) observed in all aerial roots of WT/35S::AtTCTP2-GFP scions was reminiscent of the transgenic primary roots, suggesting an important role of AtTCTP2 and its nuclear localization in midroot growth and lateral root development; this also suggests that AtTCTP2 is capable of moving into the scion and into nuclei of aerial roots, which appear to be induced by AtTCTP2 itself. It must be noted that in these roots fluorescence is not restricted to the nucleus. Instead, signal can also be observed in the cell periphery, and, at lower levels, in cytoplasm; this is much higher than the background autofluorescence of WT roots.


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)

Comparison of the protein localization pattern between adventitious and primary roots in heterografts. Mid-root region from aerial or primary roots were used to identify AtTCTP2-GFP signal through fluorescence confocal microscopy. AtTCTP2-GFP nuclear localization (N) in the mid-root region of (A,D) aerial and (B,E) transgenic primary roots was recurrent in all cases, while in the (C,F) WT primary control no nuclear signal was found (N; dashed white circles). In addition, AtTCTP2-GFP was located in lateral root primordia (LRp) both in (G) aerial roots and (H) transgenic primary roots in contrast to (I) WT primary roots. Size bars: 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 6: Comparison of the protein localization pattern between adventitious and primary roots in heterografts. Mid-root region from aerial or primary roots were used to identify AtTCTP2-GFP signal through fluorescence confocal microscopy. AtTCTP2-GFP nuclear localization (N) in the mid-root region of (A,D) aerial and (B,E) transgenic primary roots was recurrent in all cases, while in the (C,F) WT primary control no nuclear signal was found (N; dashed white circles). In addition, AtTCTP2-GFP was located in lateral root primordia (LRp) both in (G) aerial roots and (H) transgenic primary roots in contrast to (I) WT primary roots. Size bars: 50 μm.
Mentions: In addition, confocal fluorescence laser microscopy was performed to detect AtTCTP2-GFP protein signal and determine its localization as well. The AtTCTP2-GFP fluorescent signal was specifically found in the nuclei of the mid-root region in all aerial roots that emerged (Figure 6A) which is consistent with the localization pattern found in the 35S::AtTCTP2-GFP transgenic primary roots (Figure 6B; Toscano-Morales et al., submitted) and in contrast with the absence of nuclear signal in the primary mid-root region of WT stock controls (Figure 6C). Interestingly, no nuclear localization signal is predicted for AtTCTP2, or several plant TCTPs tested, using a nuclear predictor server (http://www.sbc.su.se/~maccallr/nucpred/). In a similar manner, the same localization pattern was observed in lateral root primordia both in aerial and transgenic primary root (Figures 6D,E,G,H), unlike the WT stock control (Figures 6F,I). Remarkably, the protein accumulation pattern (in cells at the base of the lateral root primordia) observed in all aerial roots of WT/35S::AtTCTP2-GFP scions was reminiscent of the transgenic primary roots, suggesting an important role of AtTCTP2 and its nuclear localization in midroot growth and lateral root development; this also suggests that AtTCTP2 is capable of moving into the scion and into nuclei of aerial roots, which appear to be induced by AtTCTP2 itself. It must be noted that in these roots fluorescence is not restricted to the nucleus. Instead, signal can also be observed in the cell periphery, and, at lower levels, in cytoplasm; this is much higher than the background autofluorescence of WT roots.

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