Research output: Contribution to journal › Article › peer-review
Cytosolic YB-1 and NSUN2 are the only proteins recognizing specific motifs present in mRNAs enriched in exosomes. / Kossinova, Olga A.; Gopanenko, Alexander V.; Tamkovich, Svetlana N. et al.
In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1865, No. 6, 01.06.2017, p. 664-673.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Cytosolic YB-1 and NSUN2 are the only proteins recognizing specific motifs present in mRNAs enriched in exosomes
AU - Kossinova, Olga A.
AU - Gopanenko, Alexander V.
AU - Tamkovich, Svetlana N.
AU - Krasheninina, Olga A.
AU - Tupikin, Alexey E.
AU - Kiseleva, Elena
AU - Yanshina, Darya D.
AU - Malygin, Alexey A.
AU - Ven'yaminova, Alia G.
AU - Kabilov, Marsel R.
AU - Karpova, Galina G.
N1 - Publisher Copyright: © 2017
PY - 2017/6/1
Y1 - 2017/6/1
N2 - Exosomes, membranous vesicles secreted by various cells, are involved in intercellular communication and carry vast repertoires of RNAs and proteins. Processes mediating RNA sorting into exosomes are currently poorly understood. Using bioinformatics approaches, three structural motifs ACCAGCCU, CAGUGAGC and UAAUCCCA have been discovered as enriched in exosomal mRNAs and long noncoding RNAs. Here, utilizing short RNA hairpins, each containing one of the motifs, in a pull-down assay of cytosolic extract of human embryonic kidney 293 (HEK293) cells, we prove that multifunctional RNA-binding protein YB-1 specifically interacts with all three motifs, whereas methyltransferase NSUN2 recognizes only the motif CAGUGAGC. RNA hairpins other than those mentioned above pull out neither YB-1 nor NSUN2. Both these proteins are found in exosomes secreted by HEK293 cells. YB-1 for all that is detected as a form having a slightly higher electrophoretic mobility than that of YB-1 associated with the above RNA hairpins, assuming changes in posttranslational modifications of the protein during its transfer from cytoplasm into exosomes. Next generation sequencing of total exosomal RNA (eRNA) reveals a large representative set of RNA species, including mRNAs containing the above-mentioned motifs. The degree of enrichment in exosomes with this kind of mRNAs strongly depends on the locations of eRNA-specific motifs within the mRNA sequences. Altogether, our findings point to YB-1 and NSUN2 as possible mediators of the process of transfer of specific mRNAs into exosomes, allowing us to speculate on an involvement of these proteins in the mRNA sorting via the recognition of the above motifs.
AB - Exosomes, membranous vesicles secreted by various cells, are involved in intercellular communication and carry vast repertoires of RNAs and proteins. Processes mediating RNA sorting into exosomes are currently poorly understood. Using bioinformatics approaches, three structural motifs ACCAGCCU, CAGUGAGC and UAAUCCCA have been discovered as enriched in exosomal mRNAs and long noncoding RNAs. Here, utilizing short RNA hairpins, each containing one of the motifs, in a pull-down assay of cytosolic extract of human embryonic kidney 293 (HEK293) cells, we prove that multifunctional RNA-binding protein YB-1 specifically interacts with all three motifs, whereas methyltransferase NSUN2 recognizes only the motif CAGUGAGC. RNA hairpins other than those mentioned above pull out neither YB-1 nor NSUN2. Both these proteins are found in exosomes secreted by HEK293 cells. YB-1 for all that is detected as a form having a slightly higher electrophoretic mobility than that of YB-1 associated with the above RNA hairpins, assuming changes in posttranslational modifications of the protein during its transfer from cytoplasm into exosomes. Next generation sequencing of total exosomal RNA (eRNA) reveals a large representative set of RNA species, including mRNAs containing the above-mentioned motifs. The degree of enrichment in exosomes with this kind of mRNAs strongly depends on the locations of eRNA-specific motifs within the mRNA sequences. Altogether, our findings point to YB-1 and NSUN2 as possible mediators of the process of transfer of specific mRNAs into exosomes, allowing us to speculate on an involvement of these proteins in the mRNA sorting via the recognition of the above motifs.
KW - Exosomal RNA-specific motifs
KW - mRNAs species enriched in exosomes
KW - NSUN2
KW - Pull-down assay of cytosolic extract
KW - RNA-protein interactions
KW - YB-1
KW - Amino Acid Sequence
KW - Humans
KW - Exosomes/metabolism
KW - Microscopy, Immunoelectron
KW - Flow Cytometry
KW - Methyltransferases/chemistry
KW - RNA, Messenger/chemistry
KW - Y-Box-Binding Protein 1/chemistry
KW - HEK293 Cells
KW - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
KW - Cytosol/metabolism
UR - http://www.scopus.com/inward/record.url?scp=85016418559&partnerID=8YFLogxK
U2 - 10.1016/j.bbapap.2017.03.010
DO - 10.1016/j.bbapap.2017.03.010
M3 - Article
C2 - 28341602
AN - SCOPUS:85016418559
VL - 1865
SP - 664
EP - 673
JO - Biochimica et Biophysica Acta - Proteins and Proteomics
JF - Biochimica et Biophysica Acta - Proteins and Proteomics
SN - 1570-9639
IS - 6
ER -
ID: 10265917