Analysis of Germline Chromatin Silencing by Double-Stranded RNA-Mediated Interference (RNAi) in &lt;I&gt;Caenorhabditis elegans&lt;B&gt;

Jedrusik, Monika A.; Schulze, Ekkehard

doi:10.1385/1-59259-741-6:035

Cited by 3 publications

(3 citation statements)

References 17 publications

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“…RNAi was induced by microinjecting dsRNA into the gonad of C. elegans and further enhanced by feeding injected worms with E. coli HT115 (DE3) expressing the specific dsRNA as described (Jedrusik and Schulze, 2004). The sir-2.2 dsRNA was microinjected into sir-2.3 ( ok444 ) mutant worms at a concentration of 2 µg/µl.…”

Section: Methodsmentioning

confidence: 99%

Mitochondrial SIRT4-type proteins in Caenorhabditis elegans and mammals interact with pyruvate carboxylase and other acetylated biotin-dependent carboxylases

Wirth¹,

Karaca²,

Wenzel³

et al. 2013

Mitochondrion

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The biological and enzymatic function of SIRT4 is largely uncharacterized. We show that the C. elegans SIR-2.2 and SIR-2.3 orthologs of SIRT4 are ubiquitously expressed, also localize to mitochondria and function during oxidative stress. Further, we identified conserved interaction with mitochondrial biotin-dependent carboxylases (PC, PCC, MCCC), key enzymes in anaplerosis and ketone body formation. The carboxylases were found acetylated on multiple lysine residues and detailed analysis of mPC suggested that one of these residues, K748ac, might regulate enzymatic activity. Nevertheless, no changes in mPC acetylation levels and enzymatic activity could be detected upon overexpression or loss of functional SIRT4.

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Section: Methodsmentioning

confidence: 99%

Mitochondrial SIRT4-type proteins in Caenorhabditis elegans and mammals interact with pyruvate carboxylase and other acetylated biotin-dependent carboxylases

Wirth¹,

Karaca²,

Wenzel³

et al. 2013

Mitochondrion

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“…Linker histone RNAi was accomplished by a combination of double-stranded-RNA injection and feeding as described earlier (9,12). For the analysis of the his-24(ok1024) mutant, two additional doublestranded RNAs were used.…”

Section: (E911) V/nt1[unc-?(n754) Let-?] (Iv;v) Mes-6(bn66) Dpy-20mentioning

confidence: 99%

Linker Histone HIS-24 (H1.1) Cytoplasmic Retention Promotes Germ Line Development and Influences Histone H3 Methylation in Caenorhabditis elegans

Jedrusik

Schulze

2007

Molecular and Cellular Biology

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RNA interference with one of the eight Caenorhabditis elegans linker histone genes triggers desilencing of a repetitive transgene and developmental defects in the hermaphrodite germ line. These characteristics are similar to the phenotype of the C. elegans Polycomb group genes mes-2, mes-3, mes-4, and mes-6 (M. A. Jedrusik and E. Schulze, Development 128:1069-1080, 2001; I. Korf, Y. Fan, and S. Strome, Development 125:2469-2478, 1998). These Polycomb group proteins contribute to germ line-specific chromatin modifications. Using a his-24 deletion mutant and an isoform-specific antibody, we characterized the role of his-24 in C. elegans germ line development. We describe an unexpected cytoplasmic retention of HIS-24 in peculiar granular structures. This phenomenon is confined to the developing germ lines of both sexes. It is strictly dependent on the activities of the chromatinmodifying genes mes-2, mes-3, mes-4, and mes-6, as well as on the C. elegans sirtuin gene sir-2.1. A temperature shift experiment with a mes-3(ts) mutant revealed that mes gene activity is required in a time window ranging from L3 to the early L4 stage before the onset of meiosis. We find that the his-24(ok1024) mutant germ line is characterized by an increased level of the activating H3K4 methylation mark concomitant with a decrease of the repressive H3K9 methylation. In the germ line of his-24(ok1024) mes-3(bn35) double mutant animals, the repressive H3K27 methylation is more reduced than in the respective mes single mutant. These observations distinguish his-24 as an unusual element in the developmental regulation of germ line chromatin structure in C. elegans.Linker histones are highly abundant chromatin proteins that bind to the elementary structural unit of the chromatin, the nucleosome. Our previous work (9) characterized the Caenorhabditis elegans linker histone variant gene his-24 (H1.1) as a gene involved in the control of hermaphrodite germ line development. Using RNA interference (RNAi), desilencing of a repetitive transgene in the germ lines of both sexes had been shown. Additionally a low-penetrant cytological gonad phenotype occurred, where the germ line substantially lacked proliferation and differentiation. This cytological phenotype was observed in hermaphrodites but not in males. The combination of both observations related his-24 to the phenotype of the C. elegans Polycomb group genes, mes-2, mes-3, mes-4, and mes-6 (18; for reviews, see references 25 and 27). The precise gonadal expression pattern of HIS-24, its general mode of action, and its specific functional relationship to the mes genes remained unclear. No germ line phenotype of a linker histone mutant has been reported for mammals so far, although the mouse linker histone complement has recently been recognized as essential for embryogenesis (6). The present view on linker histones in general describes them as highly dynamic chromatin components. They are considered to be dispensable in single-cell eukaryotes (4, 16).The C. elegans SET domain histone methyl trans...

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“…RNAi was induced by microinjecting dsRNA into the gonad of C. elegans and further enhanced by feeding injected worms with E. coli HT115 (DE3) expressing the specific dsRNA [186].…”

Section: Rna Interference (Rnai) By Microinjection and Feedingmentioning

confidence: 99%

Functional analysis of mitochondrial sirtuins in C. elegans and mammalian cells

Wirth¹

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Analysis of Germline Chromatin Silencing by Double-Stranded RNA-Mediated Interference (RNAi) in <I>Caenorhabditis elegans<B>

Cited by 3 publications

References 17 publications

Mitochondrial SIRT4-type proteins in Caenorhabditis elegans and mammals interact with pyruvate carboxylase and other acetylated biotin-dependent carboxylases

Mitochondrial SIRT4-type proteins in Caenorhabditis elegans and mammals interact with pyruvate carboxylase and other acetylated biotin-dependent carboxylases

Linker Histone HIS-24 (H1.1) Cytoplasmic Retention Promotes Germ Line Development and Influences Histone H3 Methylation in Caenorhabditis elegans

Functional analysis of mitochondrial sirtuins in C. elegans and mammalian cells

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