<i>Caenorhabditis elegans</i>
            ALG-1 antimorphic mutations uncover functions for Argonaute in microRNA guide strand selection and passenger strand disposal

Zinovyeva, Anna Y.; Veksler-Lublinsky, Isana; Vashisht, Ajay A.; Wohlschlegel, James A.; Ambros, Victor R.

doi:10.1073/pnas.1506576112

Cited by 29 publications

(77 citation statements)

References 59 publications

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“…In this manuscript we report characterizations of hrpk-1, which encodes an RNA binding protein HRPK-1. HRPK-1 was originally identified by MudPIT proteomics in ALG-1 co-precipitates [14], ( Fig 1D). Here, we confirm that HRPK-1 can co-precipitate ALG-1 in a reciprocal experiment and show that the interaction between HRPK-1 and ALG-1 is only partially RNA-dependent (Fig1E).…”

Section: Hrpk-1 Physically and Functionally Interacts With Mirna Pathmentioning

confidence: 98%

“…We have previously identified HRPK-1(F26B1.2) by immunoprecipitating C. elegans miRNAspecific Argonaute ALG-1 and analyzing the co-purified protein complexes by MudPIT proteomics [14], ( Fig 1D). To confirm this physical interaction, we performed reciprocal IP using antisera generated against HRPK-1 and probed for the presence of ALG-1.…”

Section: Alg-1 Argonaute and Hrpk-1 Co-immunoprecipitate In A Partialmentioning

confidence: 99%

“…These include the C. elegans translational repressor, a Quaking homolog, and a KH domain protein, GLD-1, among others [28], [41] and VGLN-1 [24]. Additional KH-domain proteins and other hnRNPs have been detected in large-scale mass spectrometry experiments [41], [14], [42]. The molecular mechanisms by which RNA binding proteins synergize with miRNA pathways to regulate mRNA expression are diverse and remain largely unknown.…”

Section: How Might Hrpk-1 Enhance Mirna Gene Repressive Activity?mentioning

confidence: 99%

“…The size selected RNA was then used to construct small RNA libraries using the NEXTflex Small RNA Library Prep kit v3 (Bioo Scientific) and sequenced on the Illumina HighSeq instrument at the Kansas University Genome Sequencing Core. Data analysis was performed as previously described [14]. The total mapped reads across two replicates were as follows: N2 (7,386,035), UY38 (7,679,177), and UY42 (7,540,761).…”

Section: Rna Preparation and Small Rna Sequencingmentioning

confidence: 99%

“…It remains unclear how many of them function as direct modulators of miRISC activity. Complementary to these approaches, we have previously sought to identify physical interactors of ALG-1, a miRNAspecific C. elegans Argonaute [14], hypothesizing that proteins that co-precipitate with ALG-1 include factors that modulate miRNA-induced gene repression. An RNAi-based screen of the putative ALG-1 co-factors has identified HRPK-1, a conserved homolog of human heterogeneous nuclear ribonucleoprotein K (hnRNPK), as a novel miRNA interactor that modulates activity of several binding near miR-122 target sites on target mRNAs [27].…”

Section: Introductionmentioning

confidence: 99%

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HRPK-1, a conserved KH-domain protein, modulates microRNA activity during Caenorhabditis elegans development

Zinovyeva

Veksler-Lublinsky

2019

Preprint

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microRNAs (miRNAs) are potent regulators of gene expression that function in diverse developmental and physiological processes. Argonaute proteins loaded with miRNAs form the miRNA Induced Silencing Complexes (miRISCs) that repress gene expression at the post-transcriptional level. miRISCs target genes through partial sequence complementarity between the miRNA and the target mRNA's 3' UTR. In addition to being targeted by miRNAs, these mRNAs are also extensively regulated by RNA-binding proteins (RBPs) through RNA processing, transport, stability, and translation regulation. While the degree to which RBPs and miRISCs functionally interact to regulate gene expression is likely extensive, we have only begun to unravel these functional interactions. AnRNAi-based screen of putative ALG-1 Argonaute interactors has identified a role for a conserved RNA binding protein, HRPK-1, in modulating miRNA activity during C. elegans development. Here, we report the physical and genetic interaction between HRPK-1 and ALG-1/miRNAs. Specifically, we report the genetic and molecular characterizations of hrpk-1 and its role in C. elegans development and miRNA-mediated target repression. We show that loss of hrpk-1 causes numerous developmental defects and enhances the mutant phenotypes associated with reduction of miRNA activity, including those of lsy-6, mir-35-family, and let-7-family miRNAs. In addition to hrpk-1 genetic interaction with these miRNA families, hrpk-1 is required for efficient regulation of lsy-6 target cog-1.We report that hrpk-1 may play a role in miRNA processing but is not globally required for mature miRNA biogenesis or ALG-1/AIN-1 miRISC assembly and confirm HRPK-1 ability to co-precipitate with ALG-1. We suggest that HRPK-1 may functionally interact with miRNAs on multiple levels to enhance miRNA/miRISC gene regulatory activity and present several models for its activity. Author summarymicroRNAs are small non-coding RNAs that regulate gene expression at the post-transcriptional level. The core microRNA Induced Silencing Complex (miRISC), composed of Argonaute, mature microRNA, and GW182 protein effector, assembles on the target messenger RNA and inhibits translation or leads to messenger RNA degradation. RNA binding proteins interface with miRNA pathways on multiple levels to coordinate gene expression regulation. Here, we report identification and characterization of HRPK-1, a conserved RNA binding protein, as a physical and functional interactor of miRNAs. We confirm the physical interaction between HRPK-1, an hnRNPK homolog, and Argonaute ALG-1. We report characterizations of hrpk-1 role in development and its functional interactions with multiple miRNA families. We suggest that HRPK-1 promotes miRNA activity on multiple levels in part by contributing to miRNA processing and by coordinating with miRISC at the level of target RNAs. This work contributes to our understanding of how RNA binding proteins and auxiliary miRNA cofactors may interface with miRNA pathways to modulate miRNA gene regulatory activity....

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Section: Hrpk-1 Physically and Functionally Interacts With Mirna Pathmentioning

confidence: 98%

Section: Alg-1 Argonaute and Hrpk-1 Co-immunoprecipitate In A Partialmentioning

confidence: 99%

Section: How Might Hrpk-1 Enhance Mirna Gene Repressive Activity?mentioning

confidence: 99%

Section: Rna Preparation and Small Rna Sequencingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

HRPK-1, a conserved KH-domain protein, modulates microRNA activity during Caenorhabditis elegans development

Zinovyeva

Veksler-Lublinsky

2019

Preprint

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microRNA strand selection: Unwinding the rules

2020

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microRNAs (miRNAs) play a central role in the regulation of gene expression by targeting specific mRNAs for degradation or translational repression. Each miRNA is post‐transcriptionally processed into a duplex comprising two strands. One of the two miRNA strands is selectively loaded into an Argonaute protein to form the miRNA‐Induced Silencing Complex (miRISC) in a process referred to as miRNA strand selection. The other strand is ejected from the complex and is subject to degradation. The target gene specificity of miRISC is determined by sequence complementarity between the Argonaute‐loaded miRNA strand and target mRNA. Each strand of the miRNA duplex has the capacity to be loaded into miRISC and possesses a unique seed sequence. Therefore, miRNA strand selection plays a defining role in dictating the specificity of miRISC toward its targets and provides a mechanism to alter gene expression in a switch‐like fashion. Aberrant strand selection can lead to altered gene regulation by miRISC and is observed in several human diseases including cancer. Previous and emerging data shape the rules governing miRNA strand selection and shed light on how these rules can be circumvented in various physiological and pathological contexts. This article is categorized under: RNA Processing > Processing of Small RNAs Regulatory RNAs/RNAi/Riboswitches > Regulatory RNAs Regulatory RNAs/RNAi/Riboswitches > Biogenesis of Effector Small RNAs

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Considering Context-Specific microRNAs in Ischemic Stroke with Three “W”: Where, When, and What

Chang,

Wang,

Wang

et al. 2024

Mol Neurobiol

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Caenorhabditis elegans ALG-1 antimorphic mutations uncover functions for Argonaute in microRNA guide strand selection and passenger strand disposal

Cited by 29 publications

References 59 publications

HRPK-1, a conserved KH-domain protein, modulates microRNA activity during Caenorhabditis elegans development

HRPK-1, a conserved KH-domain protein, modulates microRNA activity during Caenorhabditis elegans development

microRNA strand selection: Unwinding the rules

Considering Context-Specific microRNAs in Ischemic Stroke with Three “W”: Where, When, and What

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