Bovine Brain Ribonuclease Is the Functional Homolog of Human Ribonuclease 1

Eller, Chelcie H.; Lomax, Jo E.; Raines, Ronald T.

doi:10.1074/jbc.m114.566166

Cited by 22 publications

(44 citation statements)

References 75 publications

(76 reference statements)

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“…These data suggest that RNases can enter these MkMPs to degrade their RNA content. While it is firmly established that RNase A, RNase 1 and likely most RNases are actively endocytosed by mammalian cells (so much so that RNase A has been termed a “cell-penetrating protein” [59, 60]), the mechanism by which RNases enter MPs like these MkMPs remains unexplored.…”

Section: Resultsmentioning

confidence: 99%

How do megakaryocytic microparticles target and deliver cargo to alter the fate of hematopoietic stem cells?

Jiang

Kao

Papoutsakis

2017

Journal of Controlled Release

122

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Megakaryocytic microparticles (MkMPs), the most abundant MPs in circulation, can induce the differentiation of hematopoietic stem and progenitor cells (HSPCs) into functional megakaryocytes. This MkMP capability could be explored for applications in transfusion medicine but also for delivery of nucleic acids and other molecules to HSPCs for targeted molecular therapy. Understanding how MkMPs target, deliver cargo and alter the fate of HSPCs is important for exploring such applications. We show that MkMPs, which are distinct from Mk exosomes (MkExos), target HSPCs with high specificity since they have no effect on other ontologically or physiologically related cells, namely mesenchymal stem cells, endothelial cells or granulocytes. The outcome is also specific: only cells of the megakaryocytic lineage are generated. Observation of intact fluorescently-tagged MkMPs inside HSPCs demonstrates endocytosis as one mechanism of cargo delivery. Fluorescent labeling and scanning electron microscopy (SEM) imaging show that direct fusion of MkMPs into HSPCs is also engaged in cargo delivery. SEM imaging detailed the membrane-fusion process in four stages leading to full adsorption of MkMPs into HSPCs. Specifically, macropinocytosis and lipid raft-mediated were shown here as mechanisms of MkMP uptake by HSPC. In contrast, the ontologically related platelet-derived MPs (PMPs) cannot be taken up by HSPCs although they bind to and induce HSPC aggregation. We show that platelet-like thrombin activation is apparently responsible for the different biological effects of MkMPs versus PMPs on HSPCs. We show that HSPC uropods are the preferential site for MkMP binding, and that CD54 (ICAM-1), CD11b, CD18 and CD43, localized on HSPC uropods, are involved in MkMP binding to HSPCs. Finally, we show that MkMP RNA is largely responsible for HSPC programming into Mk differentiation.

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Section: Resultsmentioning

confidence: 99%

How do megakaryocytic microparticles target and deliver cargo to alter the fate of hematopoietic stem cells?

Jiang

Kao

Papoutsakis

2017

Journal of Controlled Release

122

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“…Brain ribonuclease is a member of the ribonuclease A superfamily of 10 to 28 kDa proteins [5–8] and was recently identified as the functional homolog of human ribonuclease-1 (RNase1) [9]. Ribonuclease A superfamily proteins are multifaceted and exhibit immuno-modulatory effects [9], antitumoral activity [10] and pro-apopototic activity [11]. Pancreatic RNase A is the better described RNase A superfamily member in cattle; this RNase is thought to function to breakdown the large amounts of RNA that accumulate in the ruminant gut [12, 13].…”

Section: Introductionmentioning

confidence: 99%

“…Pancreatic RNase A is the better described RNase A superfamily member in cattle; this RNase is thought to function to breakdown the large amounts of RNA that accumulate in the ruminant gut [12, 13]. Another well-known RNase A superfamily member, seminal ribonuclease, is produced by bovine seminal vesicles, shares 80% identity with bovine pancreatic RNase A and has cytotoxic and immunosuppressive activity needed to protect spermatozoa from the female immune system [9, 14]. The ribonuclease A superfamily member, RNase5 (also called angiogenin), has been linked to morphological changes in the bovine ovary [15] and angiogenesis [16], but whether RNase1 homologs such as BRB change during follicular development or regulate follicular atresia is unknown.…”

Section: Introductionmentioning

confidence: 99%

Changes in brain ribonuclease (BRB) messenger RNA in granulosa cells (GCs) of dominant vs subordinate ovarian follicles of cattle and the regulation of BRB gene expression in bovine GCs

Dentis

Schreiber

Gilliam

et al. 2016

Domestic Animal Endocrinology

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Brain ribonuclease (BRB) is a member of the ribonuclease A superfamily that is constitutively expressed in a range of tissues, and is the functional homolog of human ribonuclease 1. This study was designed to characterize BRB gene expression in granulosa cells (GC) during development of bovine dominant ovarian follicles, and to determine the hormonal regulation of BRB in GC. Estrous cycles of Holstein cows (n = 18) were synchronized and cows were ovariectomized on either day 3 to 4 or day 5 to 6 post-ovulation during dominant follicle growth and selection. Ovaries were collected, follicular fluid (FFL) was aspirated, and GC were collected for RNA isolation and quantitative PCR. Follicles were categorized as small (1 to 5 mm; pooled per ovary), medium (5 to 8 mm; individually collected) or large (8.1 to 17 mm; individually collected) based on surface diameter. Estradiol (E2) and progesterone (P4) levels were measured by RIA in FFL. Abundance of BRB mRNA in GC was 8.6- to 11.8-fold greater (P < 0.05) in small (n = 31), medium (n = 66) and large (n = 33) subordinate E2-inactive (FFL E2 < P4) follicles than in large (n = 16) dominant E2-active (FFL E2 > P4) follicles. In the largest 4 follicles, GC BRB mRNA abundance was negatively correlated (P < 0.01) with FFL E2 (r = −0.65) and E2/P4 ratio (r = −0.46). In Exp. 2, GC from large (8 to 22 mm diameter) and small (1 to 5 mm diameter) follicles were treated with IGF1 (0 or 30 ng/mL), and/or tumor necrosis factor α (TNFα) (0 or 30 ng/mL); IGF1 increased (P < 0.05) BRB mRNA abundance and TNFα decreased (P < 0.001) the IGF1-induced BRB mRNA abundance in large-follicle GC. In Exp. 3 to 6, E2, FSH, fibroblast growth factor 9 (FGF9), cortisol, wingless 3A (WNT3A), or Sonic hedgehog (SHH) did not affect (P > 0.10) abundance of BRB mRNA in GC; thyroxine and LH increased (P < 0.05) whereas prostaglandin E2 (PGE2) decreased (P < 0.05) BRB mRNA abundance in small-follicle GC. Treatment of small-follicle GC with recombinant human RNase1 increased (P < 0.05) GC numbers and estradiol production. In conclusion, BRB is a hormonally and developmentally regulated gene in bovine GC and may regulate estradiol production during follicular growth in cattle.

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“…33–35 This enzyme can re-enter endothelial cells by endocytosis, suggesting that the cytosol is assaulted constantly by a potent catalyst of RNA degradation. 32,36–38 By binding and inactivating RNase 1, 6 RNH1 can protect cytosolic RNA from degradation and promote cell survival.…”

mentioning

confidence: 99%

Knockout of the Ribonuclease Inhibitor Gene Leaves Human Cells Vulnerable to Secretory Ribonucleases

2016

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Ribonuclease inhibitor (RNH1) is a cytosolic protein that binds with femtomolar affinity to human ribonuclease 1 (RNase 1) and homologous secretory ribonucleases. RNH1 contains 32 cysteine residues and has been implicated as an anti-oxidant. Here, we use CRISPR–Cas9 to knockout RNH1 in HeLa cells. We find that cellular RNH1 affords marked protection from the lethal ribonucleolytic activity of RNase 1 but not from oxidants. We conclude that RNH1 protects cytosolic RNA from invading ribonucleases.

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Bovine Brain Ribonuclease Is the Functional Homolog of Human Ribonuclease 1

Cited by 22 publications

References 75 publications

How do megakaryocytic microparticles target and deliver cargo to alter the fate of hematopoietic stem cells?

How do megakaryocytic microparticles target and deliver cargo to alter the fate of hematopoietic stem cells?

Changes in brain ribonuclease (BRB) messenger RNA in granulosa cells (GCs) of dominant vs subordinate ovarian follicles of cattle and the regulation of BRB gene expression in bovine GCs

Knockout of the Ribonuclease Inhibitor Gene Leaves Human Cells Vulnerable to Secretory Ribonucleases

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