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Molecular and Cellular Biology, August 2005, p. 7107-7119, Vol. 25, No. 16
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.16.7107-7119.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

HANP1/H1T2, a Novel Histone H1-Like Protein Involved in Nuclear Formation and Sperm Fertility

Hiromitsu Tanaka,1 Naoko Iguchi,1 Ayako Isotani,2 Kouichi Kitamura,1 Yoshiro Toyama,3 Yasuhiro Matsuoka,1 Masayoshi Onishi,1 Kumiko Masai,1 Mamiko Maekawa,3 Kiyotaka Toshimori,3 Masaru Okabe,2 and Yoshitake Nishimune1*

Department of Science for Laboratory Animal Experimentation, Research Institute for Microbial Diseases,1 Genome Information Research Center, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871,2 Department of Anatomy and Developmental Biology, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan3

Received 19 February 2005/ Returned for modification 26 April 2005/ Accepted 20 May 2005

We cloned a testis-specific cDNA from mice that encodes a histone H1-like, haploid germ cell-specific nuclear protein designated HANP1/H1T2. The HANP1/H1T2 protein was specifically localized to the nuclei of murine spermatids during differentiation steps 5 to 13 but not to the nuclei of mature sperm. HANP1/H1T2 contains an arginine-serine-rich domain and an ATP/GTP binding site, and it binds to DNA, ATP, and protamine. To investigate the physiological role of HANP1/H1T2, we generated Hanp1/H1T2-disrupted mutant mice. Homozygous Hanp1/H1T2 mutant males were infertile, but females were fertile. Although a substantial number of sperm were recovered from the epididymides, their shape and function were abnormal. During sperm morphogenesis, the formation of nuclei was disturbed and protamine-1 and -2 were only weakly detectable in the nuclei. The chromatin packaging was aberrant, as demonstrated by electron microscopy and biochemical analysis. The mutant sperm exhibited deficient motility and were not competent to fertilize eggs under in vitro fertilization conditions; however, they were capable of fertilizing eggs via intracytoplasmic sperm injection that resulted in the birth of healthy progeny. Thus, we found that HANP1/H1T2 is essential for nuclear formation in functional spermatozoa and is specifically involved in the replacement of histones with protamines during spermiogenesis. At the time of submission of the manuscript, we found an independent publication by Martianov et al. (I. Martianov, S. Brancorsini, R. Catena, A. Gansmuller, N. Kotaja, M. Parvinen, P. Sassone-Corsi, and I. Davidson, Proc. Natl. Acad. Sci. USA 102:2808-2813, 2005) that reported similar results.

* Corresponding author. Mailing address: Department of Science for Laboratory Animal Experimentation, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan. Phone: 81-6-6879-8335. Fax: 81-6-6879-8339. E-mail: nishimun{at}biken.osaka-u.ac.jp.

Molecular and Cellular Biology, August 2005, p. 7107-7119, Vol. 25, No. 16
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.16.7107-7119.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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