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Haploinsufficiency of protamine-1 or -2 causes infertility in mice

Abstract

Protamines are the major DNA-binding proteins in the nucleus of sperm in most vertebrates1,2,3,4,5 and package the DNA in a volume less than 5% of a somatic cell nucleus6. Many mammals have one protamine, but a few species, including humans and mice, have two7,8,9,10,11,12,13. Here we use gene targeting to determine if the second protamine provides redundancy to an essential process, or if both protamines are necessary. We disrupted the coding sequence of one allele of either Prm1 or Prm2 in embryonic stem (ES) cells derived from 129-strain mice, and injected them into blastocysts from C57BL/6-strain mice. Male chimeras produced 129-genotype sperm with disrupted Prm1 or Prm2 alleles, but failed to sire offspring carrying the 129 genome. We also found that a decrease in the amount of either protamine disrupts nuclear formation, processing of protamine-2 and normal sperm function. Our studies show that both protamines are essential and that haploinsufficiency caused by a mutation in one allele of Prm1 or Prm2 prevents genetic transmission of both mutant and wild-type alleles.

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Figure 1: Strategy for targeted disruption of Prm1 (a) and Prm2 (b).
Figure 2: Fertility and genetic composition of sperm from chimeras produced with Prm1+/− (a) or Prm2+/− (b) ES cells.
Figure 3: Abnormalities of sperm from chimeras.
Figure 4: Analysis of basic proteins extracted from sperm.

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Acknowledgements

We thank R. Balhorn for providing antibodies and C. Rogers for help with experiments. Animal care was in accordance with U.S. Public Health Service guidelines for use of animals, and all procedures were approved in advance by the Institutional Animal Care and Use Committee of the National Institute of Environmental Health Sciences.

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Correspondence to Edward M. Eddy.

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Cho, C., Willis, W., Goulding, E. et al. Haploinsufficiency of protamine-1 or -2 causes infertility in mice. Nat Genet 28, 82–86 (2001). https://doi.org/10.1038/ng0501-82

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