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THOC5/FMIP, an mRNA export TREX complex protein, is essential for hematopoietic primitive cell survival in vivo.

Mancini A, Niemann-Seyde SC, Pankow R, El Bounkari O, Klebba-Färber S, Koch A, Jaworska E, Spooncer E, Gruber AD, Whetton AD, Tamura T - BMC Biol. (2010)

Bottom Line: Thus we considered the hematopoietic system and found that seven days after poly injection, the number of blood cells in peripheral blood decreased drastically.THOC5/Fms interacting protein is an essential element in the maintenance of hematopoiesis.Furthermore, mechanistically depletion of THOC5/Fms interacting protein causes the down-regulation of its direct interacting partner, THOC1 which may contribute to altered THO complex function and cell death.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institut fuer Biochemie, OE4310, Medizinische Hochschule Hannover, Carl-Neuberg-Str, 1, D-30623 Hannover, Germany. annalisa_mancini@hotmail.de

ABSTRACT

Background: The transcription/export complex is evolutionarily conserved from yeast to man and is required for coupled transcription elongation and nuclear export of mRNAs. FMIP(Fms interacting protein) is a member of the THO (suppressors of the transcriptional defects of hpr1delta by overexpression) complex which is a subcomplex of the transcription/export complex. THO complex (THOC) components are not essential for bulk poly (A)+ RNA export in higher eukaryotes, but for the nuclear export of subset of mRNAs, however, their exact role is still unclear.

Results: To study the role of THOC5/Fms interacting protein in vivo, we generated THOC5/Fms interacting protein knockout mice. Since these mice are embryonic lethal, we then generated interferon inducible conditional THOC5/Fms interacting protein knockout mice. After three poly injections all of the mice died within 14 days. No pathological alterations, however, were observed in liver, kidney or heart. Thus we considered the hematopoietic system and found that seven days after poly injection, the number of blood cells in peripheral blood decreased drastically. Investigation of bone marrow cells showed that these became apoptotic within seven days after poly injection. Committed myeloid progenitor cells and cells with long term reconstituting potential were lost from bone marrow within four days after poly injection. Furthermore, infusion of normal bone marrow cells rescued mice from death induced by loss of THOC5/Fms interacting protein.

Conclusion: THOC5/Fms interacting protein is an essential element in the maintenance of hematopoiesis. Furthermore, mechanistically depletion of THOC5/Fms interacting protein causes the down-regulation of its direct interacting partner, THOC1 which may contribute to altered THO complex function and cell death.

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THOC5/FMIP deletion causes reduction of spleen size and induction of cleaved caspase 3. Two hundred and fifty micrograms of poly (I:C) were injected into six-week-old Mx-cre THOC5/FMIP (flox/flox) (n = 9) or control THOC5/FMIP (flox/flox) (n = 6) mice three times at two-to-three-day intervals. (A) Weight of body, liver, spleen and heart. (B) Macrography of liver and spleen: seven days after the first poly (I:C) treatment. Mx-cre -: control mice (THOC5/FMIP (flox/flox)); Mx-cre +: THOC5/FMIP depleted mice (Mx-cre THOC5/FMIP (flox/flox)). (C, D) Seven days after the first poly (I:C) injection (3×), mouse spleens, heart, liver were fixed in formalin. Paraffin sections were stained by hematoxylin and eosin (C). Spleen and liver sections were supplied for immunohitochemical staining using cleaved caspase 3 (D). Original magnification: ×100 for C and ×200 for D.
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Figure 4: THOC5/FMIP deletion causes reduction of spleen size and induction of cleaved caspase 3. Two hundred and fifty micrograms of poly (I:C) were injected into six-week-old Mx-cre THOC5/FMIP (flox/flox) (n = 9) or control THOC5/FMIP (flox/flox) (n = 6) mice three times at two-to-three-day intervals. (A) Weight of body, liver, spleen and heart. (B) Macrography of liver and spleen: seven days after the first poly (I:C) treatment. Mx-cre -: control mice (THOC5/FMIP (flox/flox)); Mx-cre +: THOC5/FMIP depleted mice (Mx-cre THOC5/FMIP (flox/flox)). (C, D) Seven days after the first poly (I:C) injection (3×), mouse spleens, heart, liver were fixed in formalin. Paraffin sections were stained by hematoxylin and eosin (C). Spleen and liver sections were supplied for immunohitochemical staining using cleaved caspase 3 (D). Original magnification: ×100 for C and ×200 for D.

Mentions: To determine the pathology underlying gene knockout-induced death we analyzed specific organs and determined the weight of body, liver and spleen of five to six-week-old Mx-cre THOC5/FMIP (flox/flox) (n = 9) and THOC5/FMIP (flox/flox) (n = 6) mice before and after three times 250 μg poly (I:C) treatment. No significant differences were found in the weight of body or liver between poly (I:C)-treated and -untreated mice or between Mx-cre THOC5/FMIP (flox/flox) and control THOC5/FMIP(flox/flox) mice seven days after poly (I:C) injection (Figure 4A and 4B). Although there was no depression of THOC5/FMIP expression level, within seven days spleen weight of all Mx-cre THOC5/FMIP (flox/flox) mice dropped to 50% (P = 0.0002) of that in non-treated or control mice (Figure 4A). We next examined the histopathology of organs from the same mice (seven days after poly (I:C) injection), including, liver, heart, spleen and kidney.


THOC5/FMIP, an mRNA export TREX complex protein, is essential for hematopoietic primitive cell survival in vivo.

Mancini A, Niemann-Seyde SC, Pankow R, El Bounkari O, Klebba-Färber S, Koch A, Jaworska E, Spooncer E, Gruber AD, Whetton AD, Tamura T - BMC Biol. (2010)

THOC5/FMIP deletion causes reduction of spleen size and induction of cleaved caspase 3. Two hundred and fifty micrograms of poly (I:C) were injected into six-week-old Mx-cre THOC5/FMIP (flox/flox) (n = 9) or control THOC5/FMIP (flox/flox) (n = 6) mice three times at two-to-three-day intervals. (A) Weight of body, liver, spleen and heart. (B) Macrography of liver and spleen: seven days after the first poly (I:C) treatment. Mx-cre -: control mice (THOC5/FMIP (flox/flox)); Mx-cre +: THOC5/FMIP depleted mice (Mx-cre THOC5/FMIP (flox/flox)). (C, D) Seven days after the first poly (I:C) injection (3×), mouse spleens, heart, liver were fixed in formalin. Paraffin sections were stained by hematoxylin and eosin (C). Spleen and liver sections were supplied for immunohitochemical staining using cleaved caspase 3 (D). Original magnification: ×100 for C and ×200 for D.
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Related In: Results  -  Collection

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Figure 4: THOC5/FMIP deletion causes reduction of spleen size and induction of cleaved caspase 3. Two hundred and fifty micrograms of poly (I:C) were injected into six-week-old Mx-cre THOC5/FMIP (flox/flox) (n = 9) or control THOC5/FMIP (flox/flox) (n = 6) mice three times at two-to-three-day intervals. (A) Weight of body, liver, spleen and heart. (B) Macrography of liver and spleen: seven days after the first poly (I:C) treatment. Mx-cre -: control mice (THOC5/FMIP (flox/flox)); Mx-cre +: THOC5/FMIP depleted mice (Mx-cre THOC5/FMIP (flox/flox)). (C, D) Seven days after the first poly (I:C) injection (3×), mouse spleens, heart, liver were fixed in formalin. Paraffin sections were stained by hematoxylin and eosin (C). Spleen and liver sections were supplied for immunohitochemical staining using cleaved caspase 3 (D). Original magnification: ×100 for C and ×200 for D.
Mentions: To determine the pathology underlying gene knockout-induced death we analyzed specific organs and determined the weight of body, liver and spleen of five to six-week-old Mx-cre THOC5/FMIP (flox/flox) (n = 9) and THOC5/FMIP (flox/flox) (n = 6) mice before and after three times 250 μg poly (I:C) treatment. No significant differences were found in the weight of body or liver between poly (I:C)-treated and -untreated mice or between Mx-cre THOC5/FMIP (flox/flox) and control THOC5/FMIP(flox/flox) mice seven days after poly (I:C) injection (Figure 4A and 4B). Although there was no depression of THOC5/FMIP expression level, within seven days spleen weight of all Mx-cre THOC5/FMIP (flox/flox) mice dropped to 50% (P = 0.0002) of that in non-treated or control mice (Figure 4A). We next examined the histopathology of organs from the same mice (seven days after poly (I:C) injection), including, liver, heart, spleen and kidney.

Bottom Line: Thus we considered the hematopoietic system and found that seven days after poly injection, the number of blood cells in peripheral blood decreased drastically.THOC5/Fms interacting protein is an essential element in the maintenance of hematopoiesis.Furthermore, mechanistically depletion of THOC5/Fms interacting protein causes the down-regulation of its direct interacting partner, THOC1 which may contribute to altered THO complex function and cell death.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institut fuer Biochemie, OE4310, Medizinische Hochschule Hannover, Carl-Neuberg-Str, 1, D-30623 Hannover, Germany. annalisa_mancini@hotmail.de

ABSTRACT

Background: The transcription/export complex is evolutionarily conserved from yeast to man and is required for coupled transcription elongation and nuclear export of mRNAs. FMIP(Fms interacting protein) is a member of the THO (suppressors of the transcriptional defects of hpr1delta by overexpression) complex which is a subcomplex of the transcription/export complex. THO complex (THOC) components are not essential for bulk poly (A)+ RNA export in higher eukaryotes, but for the nuclear export of subset of mRNAs, however, their exact role is still unclear.

Results: To study the role of THOC5/Fms interacting protein in vivo, we generated THOC5/Fms interacting protein knockout mice. Since these mice are embryonic lethal, we then generated interferon inducible conditional THOC5/Fms interacting protein knockout mice. After three poly injections all of the mice died within 14 days. No pathological alterations, however, were observed in liver, kidney or heart. Thus we considered the hematopoietic system and found that seven days after poly injection, the number of blood cells in peripheral blood decreased drastically. Investigation of bone marrow cells showed that these became apoptotic within seven days after poly injection. Committed myeloid progenitor cells and cells with long term reconstituting potential were lost from bone marrow within four days after poly injection. Furthermore, infusion of normal bone marrow cells rescued mice from death induced by loss of THOC5/Fms interacting protein.

Conclusion: THOC5/Fms interacting protein is an essential element in the maintenance of hematopoiesis. Furthermore, mechanistically depletion of THOC5/Fms interacting protein causes the down-regulation of its direct interacting partner, THOC1 which may contribute to altered THO complex function and cell death.

Show MeSH