conserved mechanism for controlling protein dynamics at distinct chromosomal sites, thus ensuring homolog juxtaposition and CO exchange.
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We thank V. Matthews for assistance; A. Severson, A. Tartakoff,
A. Almasan, B. Li, M. Lichten, R. Pezza, and R. Kondratov for advice;
N. Hunter for communicating unpublished results; and A. Amon,
S. Ben-Aroya, D. Dawson, S. Keeney, A. Peyroche, A. Villeneuve,
and M. Zetka for strains and antibodies. Research was supported by
NIH grants R15GM099056 (G.V.B.), R01GM104007 (J.L. Y.) and
R01HD083177 (P.A.H.). Additional funding was provided by CSU’s Faculty
Research Development Program, John Vitullo’s Bridge Funding Program,
and the Center for Gene Regulation in Health and Disease (to G.V.B.).
Materials and Methods
Figs. S1 to S13
Tables S1 and S2
15 February 2016;
accepted 5 December 2016
Published online 5 January 2017
SCIENCE sciencemag.org 27 JANUAR Y 2017 • VOL 355 ISSUE 6323 411
Fig. 4. Meiosis-specific recruitment of the 26S proteasome to chromosomes is evolutionarily conserved. (A) Localization of the proteasome CP
(a5Pup2) and Zip1 on WT G0/1-arrested (t = 0 hours), leptotene (t = 2 hours),
early pachytene (t = 5 hours), and late pachytene/diplotene (t = 6 hours) nuclear
spreads. Bar 1, mm. (B) Same as (A), but for proteasome RP component Rpn12-
GFP without the sample at 2 hours. (C) CP (a5Pup2-GFP) and Zip1 localization in
spo11-yf (t = 5 hours), zip3D (t = 6 hours), and zip1D (t = 6 hours). Arrows, CP
foci associated with polycomplexes. Bar, 1 mm. (D) CP focus counts in WT, spo11-
yf, zip3D, and zip1D. Pachytene (Zip1 class III; red), meiotic divisions (blue). §For
focus scoring see (12) and fig. S11. Asterisks indicate significant differences
versus corresponding WT sample (two-tailed Wilcoxon rank sum test): **P <
0.01; *P < 0.05; n.s., P > 0.05. (E) RP focus counts in WT (see also Fig. 4D).
(F) Requirement for C. elegans a3PAS-3 CP for SC axis and central element
assembly. Region 1 (transition zone) excerpts from (i) WTand (ii) pas-3(RNAi)
knockdown animals stained with HTP-3 and SYP-1. Arrows indicate HTP-3/
SYP-1 coaggregates (see also fig. S12). Bar, 4 mm. (G) Squashed pachytene
nucleus stained with antibodies against CP20S and SYP-1. Overlap is similar in
≥99% of nuclei (n > 100 nuclei) from 20 germ lines (see also fig. S13B). Bar,
2 mm. (H) Proteasome and SYCP3 localization along surface-spread mouse
spermatocytes at the leptotene (n = 5), zygotene (n = 34), pachytene (n =
115), or diplotene stages (n = 47). Bars, 5 mm. Insets show magnified views
of boxed areas (see also fig. S13, C and D). (I) Model for proteasome
functions in homolog pairing, synapsis (SC), and crossover (CO) formation
(see text for details).
resubmitted 12 September 2016
Accepted 5 December 2016