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Exosomes in human semen carry a distinctive repertoire of small non-coding RNAs with potential regulatory functions.

Vojtech L, Woo S, Hughes S, Levy C, Ballweber L, Sauteraud RP, Strobl J, Westerberg K, Gottardo R, Tewari M, Hladik F - Nucleic Acids Res. (2014)

Bottom Line: We found that seminal exosome (SE) preparations contain a substantial amount of RNA from 20 to 100 nucleotides (nts) in length.We found various classes of small non-coding RNA, including microRNA (21.7% of the RNA in the 20-40 nt fraction) as well as abundant Y RNAs and tRNAs present in both fractions.Specific RNAs were consistently present in all donors.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, University of Washington, Seattle, USA luciav@uw.edu.

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Related in: MedlinePlus

SE carry Y RNA and the binding partner Ro protein. (A) Bar chart of the distribution of Y RNA in larger or smaller libraries by type. Percent reads mapping to each type are averaged across the six donors. (B) Western blot using anti-Ro antibody against SE lysate and exosome-depleted seminal plasma. SE lysates and plasma from four different individual SE samples were tested. HeLa cell lysate served as a positive control.
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Figure 4: SE carry Y RNA and the binding partner Ro protein. (A) Bar chart of the distribution of Y RNA in larger or smaller libraries by type. Percent reads mapping to each type are averaged across the six donors. (B) Western blot using anti-Ro antibody against SE lysate and exosome-depleted seminal plasma. SE lysates and plasma from four different individual SE samples were tested. HeLa cell lysate served as a positive control.

Mentions: Y RNAs are evolutionarily conserved small RNAs (83–112 nt in humans) that fold into characteristic stem-loop structures similar to tRNAs. Y RNAs are known as components of the autoantigen ribonucleoprotein complexes targeted by antibodies from patients with Sjogren's syndrome and systemic lupus erythematosus (SLE) (62). Their physiological functions are poorly characterized, though they are reported to be involved in chromosomal DNA replication, in the response to UV damage, and in regulating small RNA biogenesis (63–69). Humans have four types of Y RNAs: hY1, hY3, hY4 and hY5, all of which are present in SE (Figure 4A). Sequencing reads demonstrate the presence of both full-length or ≥50 nt fragments (in the larger size-fractionated libraries) and smaller fragmented Y RNA forms (<50 nts, in the smaller size-fractionated libraries), but the relative distribution of Y RNA types differed between the two libraries. The majority of Y RNA in both libraries was hY4. The larger library contained about 30% hY5, while the smaller library contained almost no hY5. Both libraries contained small amounts of hY1 and hY3 (Figure 4A). The ratios of larger to smaller fragmented Y RNA read counts for the four Y RNA types were extremely consistent between the six donors (Table 5). Many Y RNAs in both size fractions mapped to genomic sequences annotated as pseudogenes and predicted genes (Supplemental File 2). For all four Y RNA types, smaller fragment lengths consistently centered around 30–33 nt (Supplemental Figure S1A). For hY1, hY3 and hY5, >95% of the smaller fragments mapped to the 5’ ends of Y RNA, while for hY4 ∼80% of the smaller fragments mapped to the 5’ end and 20% mapped to the 3’ end (Supplemental Figure S1B). These data indicate that each Y RNA type is loaded into SE in a distinctive manner, implying a selective process.


Exosomes in human semen carry a distinctive repertoire of small non-coding RNAs with potential regulatory functions.

Vojtech L, Woo S, Hughes S, Levy C, Ballweber L, Sauteraud RP, Strobl J, Westerberg K, Gottardo R, Tewari M, Hladik F - Nucleic Acids Res. (2014)

SE carry Y RNA and the binding partner Ro protein. (A) Bar chart of the distribution of Y RNA in larger or smaller libraries by type. Percent reads mapping to each type are averaged across the six donors. (B) Western blot using anti-Ro antibody against SE lysate and exosome-depleted seminal plasma. SE lysates and plasma from four different individual SE samples were tested. HeLa cell lysate served as a positive control.
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Related In: Results  -  Collection

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Figure 4: SE carry Y RNA and the binding partner Ro protein. (A) Bar chart of the distribution of Y RNA in larger or smaller libraries by type. Percent reads mapping to each type are averaged across the six donors. (B) Western blot using anti-Ro antibody against SE lysate and exosome-depleted seminal plasma. SE lysates and plasma from four different individual SE samples were tested. HeLa cell lysate served as a positive control.
Mentions: Y RNAs are evolutionarily conserved small RNAs (83–112 nt in humans) that fold into characteristic stem-loop structures similar to tRNAs. Y RNAs are known as components of the autoantigen ribonucleoprotein complexes targeted by antibodies from patients with Sjogren's syndrome and systemic lupus erythematosus (SLE) (62). Their physiological functions are poorly characterized, though they are reported to be involved in chromosomal DNA replication, in the response to UV damage, and in regulating small RNA biogenesis (63–69). Humans have four types of Y RNAs: hY1, hY3, hY4 and hY5, all of which are present in SE (Figure 4A). Sequencing reads demonstrate the presence of both full-length or ≥50 nt fragments (in the larger size-fractionated libraries) and smaller fragmented Y RNA forms (<50 nts, in the smaller size-fractionated libraries), but the relative distribution of Y RNA types differed between the two libraries. The majority of Y RNA in both libraries was hY4. The larger library contained about 30% hY5, while the smaller library contained almost no hY5. Both libraries contained small amounts of hY1 and hY3 (Figure 4A). The ratios of larger to smaller fragmented Y RNA read counts for the four Y RNA types were extremely consistent between the six donors (Table 5). Many Y RNAs in both size fractions mapped to genomic sequences annotated as pseudogenes and predicted genes (Supplemental File 2). For all four Y RNA types, smaller fragment lengths consistently centered around 30–33 nt (Supplemental Figure S1A). For hY1, hY3 and hY5, >95% of the smaller fragments mapped to the 5’ ends of Y RNA, while for hY4 ∼80% of the smaller fragments mapped to the 5’ end and 20% mapped to the 3’ end (Supplemental Figure S1B). These data indicate that each Y RNA type is loaded into SE in a distinctive manner, implying a selective process.

Bottom Line: We found that seminal exosome (SE) preparations contain a substantial amount of RNA from 20 to 100 nucleotides (nts) in length.We found various classes of small non-coding RNA, including microRNA (21.7% of the RNA in the 20-40 nt fraction) as well as abundant Y RNAs and tRNAs present in both fractions.Specific RNAs were consistently present in all donors.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, University of Washington, Seattle, USA luciav@uw.edu.

Show MeSH
Related in: MedlinePlus