Limits...
Differential RNA-binding activity of the hnRNP G protein correlated with the sex genotype in the amphibian oocyte.

Kanhoush R, Praseuth D, Perrin C, Chardard D, Vinh J, Penrad-Mobayed M - Nucleic Acids Res. (2011)

Bottom Line: A proteomic approach has enabled the identification of an orthologue of the splicing factor hnRNP G in the amphibians Xenopus tropicalis, Ambystoma mexicanum, Notophthalmus viridescens and Pleurodeles walt, which shows a specific RNA-binding affinity similar to that of the human hnRN G protein.In situ hybridization to lampbrush chromosomes of P. waltl revealed the presence of a family of hnRNP G genes, which were mapped on the Z and W chromosomes and one autosome.This indicates that the isoforms identified in this study are possibly encoded by a gene family linked to the evolution of sex chromosomes similarly to the hnRNP G/RBMX gene family in mammals.

View Article: PubMed Central - PubMed

Affiliation: Institut Jacques Monod, UMR 7592, CNRS and Université Paris-Diderot, Museum National d'Histoire Naturelle, U 565, USM 503, UMR 5153, INSERM and CNRS, Paris, France.

ABSTRACT
A proteomic approach has enabled the identification of an orthologue of the splicing factor hnRNP G in the amphibians Xenopus tropicalis, Ambystoma mexicanum, Notophthalmus viridescens and Pleurodeles walt, which shows a specific RNA-binding affinity similar to that of the human hnRN G protein. Three isoforms of this protein with a differential binding affinity for a specific RNA probe were identified in the P. walt oocyte. In situ hybridization to lampbrush chromosomes of P. waltl revealed the presence of a family of hnRNP G genes, which were mapped on the Z and W chromosomes and one autosome. This indicates that the isoforms identified in this study are possibly encoded by a gene family linked to the evolution of sex chromosomes similarly to the hnRNP G/RBMX gene family in mammals.

Show MeSH
Pleurodeles waltl 42 kDa polypeptide(s) bind the WEc RNA specifically in the ZW and WW GVs. (A) Restriction map of the clone λ130 from which the RNA probes were derived by in vitro transcription. (B) NWA of 32P-labelled RNA probes with nuclear proteins from ZZ, ZW and WW GVs. Nuclear proteins were transferred to nitrocellulose membrane after electrophoresis in mono-dimensional polyacrylamide gels and incubated with the RNA-probes. Note that nuclear protein(s) in the 42 kDa range specifically bound to the WEc and PB6 RNA probes in ZW and WW karyotypes and not in ZZ karyotypes (arrow and box). No interaction could be detected for these proteins with the other RNA probes tested.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3105392&req=5

Figure 1: Pleurodeles waltl 42 kDa polypeptide(s) bind the WEc RNA specifically in the ZW and WW GVs. (A) Restriction map of the clone λ130 from which the RNA probes were derived by in vitro transcription. (B) NWA of 32P-labelled RNA probes with nuclear proteins from ZZ, ZW and WW GVs. Nuclear proteins were transferred to nitrocellulose membrane after electrophoresis in mono-dimensional polyacrylamide gels and incubated with the RNA-probes. Note that nuclear protein(s) in the 42 kDa range specifically bound to the WEc and PB6 RNA probes in ZW and WW karyotypes and not in ZZ karyotypes (arrow and box). No interaction could be detected for these proteins with the other RNA probes tested.

Mentions: The WEc RNA probe was used in NWA to identify nuclear proteins mediating its binding to the W chromosome. Proteins extracts of GVs of ZZ, ZW and WW oocytes were separated by mono-dimensional electrophoresis in polyacrylamide gels and transferred to nitrocellulose membranes. Membranes were probed with 32P-labelled RNA probes. In parallel to the WEc RNA, other RNA probes, which did not label the W chromosome lampbrush loops, were used as controls (Figure 1). In repeated NWA experiments, the WEc RNA probe showed a very similar pattern of binding proteins for the three karyotypes except in the 42 kDa range where strong binding signals were present for the ZW and WW GVs while no signal was observed for the ZZ GVs. The PB6 RNA probe showed the same differential interaction with the 42 kDa polypeptide(s) for the ZW and WW GVs although the binding signals were weaker than the ones observed with the WEc RNA. The other RNA probes (PP6, PB7 and BP7) did not bind to these polypeptides although they bound to other polypeptides for all three oocyte karyotypes. These data provided evidence for sequence-specific RNA-binding polypeptide(s) whose expression in the oocyte could be correlated with the presence of the W chromosome in its karyotype.Figure 1.


Differential RNA-binding activity of the hnRNP G protein correlated with the sex genotype in the amphibian oocyte.

Kanhoush R, Praseuth D, Perrin C, Chardard D, Vinh J, Penrad-Mobayed M - Nucleic Acids Res. (2011)

Pleurodeles waltl 42 kDa polypeptide(s) bind the WEc RNA specifically in the ZW and WW GVs. (A) Restriction map of the clone λ130 from which the RNA probes were derived by in vitro transcription. (B) NWA of 32P-labelled RNA probes with nuclear proteins from ZZ, ZW and WW GVs. Nuclear proteins were transferred to nitrocellulose membrane after electrophoresis in mono-dimensional polyacrylamide gels and incubated with the RNA-probes. Note that nuclear protein(s) in the 42 kDa range specifically bound to the WEc and PB6 RNA probes in ZW and WW karyotypes and not in ZZ karyotypes (arrow and box). No interaction could be detected for these proteins with the other RNA probes tested.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3105392&req=5

Figure 1: Pleurodeles waltl 42 kDa polypeptide(s) bind the WEc RNA specifically in the ZW and WW GVs. (A) Restriction map of the clone λ130 from which the RNA probes were derived by in vitro transcription. (B) NWA of 32P-labelled RNA probes with nuclear proteins from ZZ, ZW and WW GVs. Nuclear proteins were transferred to nitrocellulose membrane after electrophoresis in mono-dimensional polyacrylamide gels and incubated with the RNA-probes. Note that nuclear protein(s) in the 42 kDa range specifically bound to the WEc and PB6 RNA probes in ZW and WW karyotypes and not in ZZ karyotypes (arrow and box). No interaction could be detected for these proteins with the other RNA probes tested.
Mentions: The WEc RNA probe was used in NWA to identify nuclear proteins mediating its binding to the W chromosome. Proteins extracts of GVs of ZZ, ZW and WW oocytes were separated by mono-dimensional electrophoresis in polyacrylamide gels and transferred to nitrocellulose membranes. Membranes were probed with 32P-labelled RNA probes. In parallel to the WEc RNA, other RNA probes, which did not label the W chromosome lampbrush loops, were used as controls (Figure 1). In repeated NWA experiments, the WEc RNA probe showed a very similar pattern of binding proteins for the three karyotypes except in the 42 kDa range where strong binding signals were present for the ZW and WW GVs while no signal was observed for the ZZ GVs. The PB6 RNA probe showed the same differential interaction with the 42 kDa polypeptide(s) for the ZW and WW GVs although the binding signals were weaker than the ones observed with the WEc RNA. The other RNA probes (PP6, PB7 and BP7) did not bind to these polypeptides although they bound to other polypeptides for all three oocyte karyotypes. These data provided evidence for sequence-specific RNA-binding polypeptide(s) whose expression in the oocyte could be correlated with the presence of the W chromosome in its karyotype.Figure 1.

Bottom Line: A proteomic approach has enabled the identification of an orthologue of the splicing factor hnRNP G in the amphibians Xenopus tropicalis, Ambystoma mexicanum, Notophthalmus viridescens and Pleurodeles walt, which shows a specific RNA-binding affinity similar to that of the human hnRN G protein.In situ hybridization to lampbrush chromosomes of P. waltl revealed the presence of a family of hnRNP G genes, which were mapped on the Z and W chromosomes and one autosome.This indicates that the isoforms identified in this study are possibly encoded by a gene family linked to the evolution of sex chromosomes similarly to the hnRNP G/RBMX gene family in mammals.

View Article: PubMed Central - PubMed

Affiliation: Institut Jacques Monod, UMR 7592, CNRS and Université Paris-Diderot, Museum National d'Histoire Naturelle, U 565, USM 503, UMR 5153, INSERM and CNRS, Paris, France.

ABSTRACT
A proteomic approach has enabled the identification of an orthologue of the splicing factor hnRNP G in the amphibians Xenopus tropicalis, Ambystoma mexicanum, Notophthalmus viridescens and Pleurodeles walt, which shows a specific RNA-binding affinity similar to that of the human hnRN G protein. Three isoforms of this protein with a differential binding affinity for a specific RNA probe were identified in the P. walt oocyte. In situ hybridization to lampbrush chromosomes of P. waltl revealed the presence of a family of hnRNP G genes, which were mapped on the Z and W chromosomes and one autosome. This indicates that the isoforms identified in this study are possibly encoded by a gene family linked to the evolution of sex chromosomes similarly to the hnRNP G/RBMX gene family in mammals.

Show MeSH