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Antagonistic roles of ubiquitin ligase HEI10 and SUMO ligase RNF212 regulate meiotic recombination.

Qiao H, Prasada Rao HB, Yang Y, Fong JH, Cloutier JM, Deacon DC, Nagel KE, Swartz RK, Strong E, Holloway JK, Cohen PE, Schimenti J, Ward J, Hunter N - Nat. Genet. (2014)

Bottom Line: Here we show that the ubiquitin ligase HEI10 (also called CCNB1IP1) is essential for this crossover/non-crossover differentiation process.As with RNF212, dosage sensitivity for HEI10 indicates that it is a limiting factor for crossing over.We suggest that SUMO and ubiquitin have antagonistic roles during meiotic recombination that are balanced to effect differential stabilization of recombination factors at crossover and non-crossover sites.

View Article: PubMed Central - PubMed

Affiliation: 1] Howard Hughes Medical Institute, University of California, Davis, Davis, California, USA. [2] Department of Microbiology & Molecular Genetics, University of California, Davis, Davis, California, USA.

ABSTRACT
Crossover recombination facilitates the accurate segregation of homologous chromosomes during meiosis. In mammals, poorly characterized regulatory processes ensure that every pair of chromosomes obtains at least one crossover, even though most recombination sites yield non-crossovers. Designation of crossovers involves selective localization of the SUMO ligase RNF212 to a minority of recombination sites, where it stabilizes pertinent factors such as MutSγ (ref. 4). Here we show that the ubiquitin ligase HEI10 (also called CCNB1IP1) is essential for this crossover/non-crossover differentiation process. In HEI10-deficient mice, RNF212 localizes to most recombination sites, and dissociation of both RNF212 and MutSγ from chromosomes is blocked. Consequently, recombination is impeded, and crossing over fails. In wild-type mice, HEI10 accumulates at designated crossover sites, suggesting that it also has a late role in implementing crossing over. As with RNF212, dosage sensitivity for HEI10 indicates that it is a limiting factor for crossing over. We suggest that SUMO and ubiquitin have antagonistic roles during meiotic recombination that are balanced to effect differential stabilization of recombination factors at crossover and non-crossover sites.

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Dosage sensitivity of the HEI10 crossover function. (a,b) Mid-pachytene nuclei from (a) wild-type (Hei10+/+) and (b) Hei10+/mei4 heterozygous mice immunostained for HEI10 (green) and SYCP3 (red). The arrows in b highlight chromosomes that lack HEI10 foci. (c) Numbers of HEI10 foci (± s.d.) per nucleus in mid-pachytene cells (23 wild-type and 33 Hei10+/mei4 nuclei). (d,e) Mid-pachytene nuclei from (d) wild-type and (e) Hei10+/mei4 heterozygous mice immunostained for MLH1 and SYCP3. The arrows in e highlight chromosomes that lack MLH1 foci. (f) Numbers of MLH1 foci per nucleus (± s.d.) in mid-pachytene cells (20 wild-type and 23 Hei10+/mei4 nuclei). (g, h) Chromosome spreads of diakinesis/metaphase I spermatocytes from (g) wild-type and (h) Hei10+/mei4 heterozygous mice stained with DAPI. (i) Numbers of chiasmata per nucleus (± s.d.) in diakinesis/metaphase I spermatocytes (25 wild-type and 29 Hei10+/mei4 nuclei). (j–l) TUNEL stained testis sections from (j) Hei10+/+, (k) Hei10+/mei4 and (l) Hei10mei4mei4 animals. (m) Quantification of TUNEL-positive apoptotic cells (± s.d.) in spermatocyte sections.Scale bars, 10 μm in a,b,d,e,g,h and 100 μm in j,k,l.
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Figure 6: Dosage sensitivity of the HEI10 crossover function. (a,b) Mid-pachytene nuclei from (a) wild-type (Hei10+/+) and (b) Hei10+/mei4 heterozygous mice immunostained for HEI10 (green) and SYCP3 (red). The arrows in b highlight chromosomes that lack HEI10 foci. (c) Numbers of HEI10 foci (± s.d.) per nucleus in mid-pachytene cells (23 wild-type and 33 Hei10+/mei4 nuclei). (d,e) Mid-pachytene nuclei from (d) wild-type and (e) Hei10+/mei4 heterozygous mice immunostained for MLH1 and SYCP3. The arrows in e highlight chromosomes that lack MLH1 foci. (f) Numbers of MLH1 foci per nucleus (± s.d.) in mid-pachytene cells (20 wild-type and 23 Hei10+/mei4 nuclei). (g, h) Chromosome spreads of diakinesis/metaphase I spermatocytes from (g) wild-type and (h) Hei10+/mei4 heterozygous mice stained with DAPI. (i) Numbers of chiasmata per nucleus (± s.d.) in diakinesis/metaphase I spermatocytes (25 wild-type and 29 Hei10+/mei4 nuclei). (j–l) TUNEL stained testis sections from (j) Hei10+/+, (k) Hei10+/mei4 and (l) Hei10mei4mei4 animals. (m) Quantification of TUNEL-positive apoptotic cells (± s.d.) in spermatocyte sections.Scale bars, 10 μm in a,b,d,e,g,h and 100 μm in j,k,l.

Mentions: Recently, recombination rate in humans has been associated with a variant in the untranslated 5′ region of HEI10/CNNB1IP1 (A. Kong, personal communication), which has the potential to alter expression level and suggests that Hei10 may be a dosage-sensitive regulator of crossing over. To determine whether the crossover function of mouse HEI10 is dosage sensitive, we analyzed spermatocytes from Hei10+/mei4 heterozygotes (Fig. 6). Indeed, significant decreases in the numbers HEI10 foci (20.6%), MLH1 foci (13.5%) and chiasmata (10%) were detected (P=0.0003, < 0.0001 and 0.0088, respectively; Mann-Whitney test). In spermatocytes, homologs not tethered by crossovers are detected by the spindle checkpoint, which triggers apoptosis24. Consistent with the reduced crossing-over seen in Hei10 +/mei4 heterozygotes, we detected a significant increase in apoptotic cells (TUNEL positive) in testes sections from Hei10+/mei4 heterozygotes (Fig. 6j–m and Supplementary Fig. 6; P < 0.0001 Mann-Whitney test).


Antagonistic roles of ubiquitin ligase HEI10 and SUMO ligase RNF212 regulate meiotic recombination.

Qiao H, Prasada Rao HB, Yang Y, Fong JH, Cloutier JM, Deacon DC, Nagel KE, Swartz RK, Strong E, Holloway JK, Cohen PE, Schimenti J, Ward J, Hunter N - Nat. Genet. (2014)

Dosage sensitivity of the HEI10 crossover function. (a,b) Mid-pachytene nuclei from (a) wild-type (Hei10+/+) and (b) Hei10+/mei4 heterozygous mice immunostained for HEI10 (green) and SYCP3 (red). The arrows in b highlight chromosomes that lack HEI10 foci. (c) Numbers of HEI10 foci (± s.d.) per nucleus in mid-pachytene cells (23 wild-type and 33 Hei10+/mei4 nuclei). (d,e) Mid-pachytene nuclei from (d) wild-type and (e) Hei10+/mei4 heterozygous mice immunostained for MLH1 and SYCP3. The arrows in e highlight chromosomes that lack MLH1 foci. (f) Numbers of MLH1 foci per nucleus (± s.d.) in mid-pachytene cells (20 wild-type and 23 Hei10+/mei4 nuclei). (g, h) Chromosome spreads of diakinesis/metaphase I spermatocytes from (g) wild-type and (h) Hei10+/mei4 heterozygous mice stained with DAPI. (i) Numbers of chiasmata per nucleus (± s.d.) in diakinesis/metaphase I spermatocytes (25 wild-type and 29 Hei10+/mei4 nuclei). (j–l) TUNEL stained testis sections from (j) Hei10+/+, (k) Hei10+/mei4 and (l) Hei10mei4mei4 animals. (m) Quantification of TUNEL-positive apoptotic cells (± s.d.) in spermatocyte sections.Scale bars, 10 μm in a,b,d,e,g,h and 100 μm in j,k,l.
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Figure 6: Dosage sensitivity of the HEI10 crossover function. (a,b) Mid-pachytene nuclei from (a) wild-type (Hei10+/+) and (b) Hei10+/mei4 heterozygous mice immunostained for HEI10 (green) and SYCP3 (red). The arrows in b highlight chromosomes that lack HEI10 foci. (c) Numbers of HEI10 foci (± s.d.) per nucleus in mid-pachytene cells (23 wild-type and 33 Hei10+/mei4 nuclei). (d,e) Mid-pachytene nuclei from (d) wild-type and (e) Hei10+/mei4 heterozygous mice immunostained for MLH1 and SYCP3. The arrows in e highlight chromosomes that lack MLH1 foci. (f) Numbers of MLH1 foci per nucleus (± s.d.) in mid-pachytene cells (20 wild-type and 23 Hei10+/mei4 nuclei). (g, h) Chromosome spreads of diakinesis/metaphase I spermatocytes from (g) wild-type and (h) Hei10+/mei4 heterozygous mice stained with DAPI. (i) Numbers of chiasmata per nucleus (± s.d.) in diakinesis/metaphase I spermatocytes (25 wild-type and 29 Hei10+/mei4 nuclei). (j–l) TUNEL stained testis sections from (j) Hei10+/+, (k) Hei10+/mei4 and (l) Hei10mei4mei4 animals. (m) Quantification of TUNEL-positive apoptotic cells (± s.d.) in spermatocyte sections.Scale bars, 10 μm in a,b,d,e,g,h and 100 μm in j,k,l.
Mentions: Recently, recombination rate in humans has been associated with a variant in the untranslated 5′ region of HEI10/CNNB1IP1 (A. Kong, personal communication), which has the potential to alter expression level and suggests that Hei10 may be a dosage-sensitive regulator of crossing over. To determine whether the crossover function of mouse HEI10 is dosage sensitive, we analyzed spermatocytes from Hei10+/mei4 heterozygotes (Fig. 6). Indeed, significant decreases in the numbers HEI10 foci (20.6%), MLH1 foci (13.5%) and chiasmata (10%) were detected (P=0.0003, < 0.0001 and 0.0088, respectively; Mann-Whitney test). In spermatocytes, homologs not tethered by crossovers are detected by the spindle checkpoint, which triggers apoptosis24. Consistent with the reduced crossing-over seen in Hei10 +/mei4 heterozygotes, we detected a significant increase in apoptotic cells (TUNEL positive) in testes sections from Hei10+/mei4 heterozygotes (Fig. 6j–m and Supplementary Fig. 6; P < 0.0001 Mann-Whitney test).

Bottom Line: Here we show that the ubiquitin ligase HEI10 (also called CCNB1IP1) is essential for this crossover/non-crossover differentiation process.As with RNF212, dosage sensitivity for HEI10 indicates that it is a limiting factor for crossing over.We suggest that SUMO and ubiquitin have antagonistic roles during meiotic recombination that are balanced to effect differential stabilization of recombination factors at crossover and non-crossover sites.

View Article: PubMed Central - PubMed

Affiliation: 1] Howard Hughes Medical Institute, University of California, Davis, Davis, California, USA. [2] Department of Microbiology & Molecular Genetics, University of California, Davis, Davis, California, USA.

ABSTRACT
Crossover recombination facilitates the accurate segregation of homologous chromosomes during meiosis. In mammals, poorly characterized regulatory processes ensure that every pair of chromosomes obtains at least one crossover, even though most recombination sites yield non-crossovers. Designation of crossovers involves selective localization of the SUMO ligase RNF212 to a minority of recombination sites, where it stabilizes pertinent factors such as MutSγ (ref. 4). Here we show that the ubiquitin ligase HEI10 (also called CCNB1IP1) is essential for this crossover/non-crossover differentiation process. In HEI10-deficient mice, RNF212 localizes to most recombination sites, and dissociation of both RNF212 and MutSγ from chromosomes is blocked. Consequently, recombination is impeded, and crossing over fails. In wild-type mice, HEI10 accumulates at designated crossover sites, suggesting that it also has a late role in implementing crossing over. As with RNF212, dosage sensitivity for HEI10 indicates that it is a limiting factor for crossing over. We suggest that SUMO and ubiquitin have antagonistic roles during meiotic recombination that are balanced to effect differential stabilization of recombination factors at crossover and non-crossover sites.

Show MeSH
Related in: MedlinePlus