Molecular Biology

, Volume 52, Issue 5, pp 707–714 | Cite as

SIVA1 Regulates the Stability of Single-Stranded DNA-Binding Protein 3 Isoforms

  • Z. YinEmail author
  • K. Zhang
  • X. Peng
  • Z. Jiang
  • W. Yuan
  • Y. Wang
  • Y. Li
  • X. Ye
  • Y. Dong
  • Y. Wan
  • B. Ni
  • P. Zhu
  • X. FanEmail author
  • X. WuEmail author
  • X. MoEmail author


The assembly of LIM-homeodomain (LIM-HD) transcriptional complex plays important roles in early neuronal development. The stability of LIM-HD is controlled by single-strand binding protein 3 (SSBP3) via a cascade mechanism protecting it from proteasomal degradation. The expression level of SSBP3 has to be precisely regulated. Although a decrease of SSBP3 level is associated with several diseases, the mechanism of SSBP3 downregulation and whether SSBP3 itself is subject to proteasomal degradation remain largely unknown. Two strongly conserved transcripts of the SSBP3 gene, SSBP3a and SSBP3c, were cloned from a human brain cDNA library. By RT-PCR, we show that Ssbp3c is continuously expressed in both embryonic and adult mouse brain, whereas Ssbp3a is restricted to embryonic brain tissue. By co-IP and GST pulldown assays, we identified SIVA1 as a novel SSBP3-binding factor. In a ubiquitination assay, we show that SIVA1 enhances the ubiquitination of SSBP3 and regulates its abundance. Our findings reveal the proteasomal degradation of SSBP3 for the first time and provide a rationale for an SIVA1-SSBP3-dependent mechanism for the disassembly of LIM-HD multiprotein complexes.


SSBP3 SIVA1 alternative splicing protein interaction ubiquitination 



This study was supported by National Natural Science Foundation of China (81400304, 81801392, 81700338, 81470449, 81470377, 81670290, 31572349, 81370451, 81670288, 81270156, 81270291), Hunan Provincial National Natural Science Foundation of China (2018JJ2666, 2015JJ3087) and the Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province 2013-448-6, the Scientific Research Fund of Hunan Provincial Education Department (no. 14A093), and China Postdoctoral Science Foundation (2018M630903).


  1. 1.
    Li W., Bengtson M.H., Ulbrich A., Matsuda A., Reddy V.A., Orth A., Chanda S.K., Batalov S., Joazeiro C.A. 2008. Genome-wide and functional annotation of human E3 ubiquitin ligases identifies MULAN, a mitochondrial E3 that regulates the organelle’s dynamics and signaling. PLoS One. 3 (1), e1487.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Bayarsaihan D., Soto R.J., Lukens L.N. 1998. Cloning and characterization of a novel sequence-specific single-stranded-DNA-binding protein. Biochem. J. 331 (2), 447‒452.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Chen L., Segal D., Hukriede N.A., Podtelejnikov A.V., Bayarsaihan D., Kennison J.A., Ogryzko V.V., Dawid I.B., Westphal H. 2002. Ssdp proteins interact with the LIM-domain-binding protein Ldb1 to regulate development. Proc. Natl. Acad. Sci. U. S. A. 99, 14 320‒14 325.CrossRefGoogle Scholar
  4. 4.
    van Meyel D.J., Thomas J.B., Agulnick A.D. 2003. Ssdp proteins bind to LIM-interacting co-factors and regulate the activity of LIM-homeodomain protein complexes in vivo. Development. 130, 1915‒1925.CrossRefPubMedGoogle Scholar
  5. 5.
    Nishioka N., Nagano S., Nakayama R., Kiyonari H., Ijiri T., Taniguchi K., Shawlot W., Hayashizaki Y., Westphal H., Behringer R.R., Matsuda Y., Sakoda S., Kondoh H., Sasaki H. 2005. Ssdp1 regulates head morphogenesis of mouse embryos by activating the Lim1–Ldb1 complex. Development. 132, 2535‒2546.CrossRefPubMedGoogle Scholar
  6. 6.
    Enkhmandakh B., Makeyev A.V., Bayarsaihan D. 2006. The role of the proline-rich domain of Ssdp1 in the modular architecture of the vertebrate head organizer. Proc. Natl. Acad. Sci. U. S. A. 103, 11 631‒11 636.CrossRefGoogle Scholar
  7. 7.
    Gungor C., Taniguchi-Ishigaki N., Ma H., Drung A., Tursun B., Ostendorff H.P., Bossenz M., Becker C.G., Becker T., Bach I. 2007. Proteasomal selection of multiprotein complexes recruited by LIM homeodomain transcription factors. Proc. Natl. Acad. Sci. U. S. A. 104, 15 000‒15 005.CrossRefGoogle Scholar
  8. 8.
    Hiratani I., Yamamoto N., Mochizuki T., Ohmori S.Y., Taira M. 2003. Selective degradation of excess Ldb1 by Rnf12/RLIM confers proper Ldb1 expression levels and Xlim-1/Ldb1 stoichiometry in Xenopus organizer functions. Development. 130, 4161‒4175.CrossRefPubMedGoogle Scholar
  9. 9.
    Ostendorff H.P., Peirano R.I., Peters M.A., Schluter A., Bossenz M., Scheffner M., Bach I. 2002. Ubiquitination-dependent cofactor exchange on LIM homeodomain transcription factors. Nature. 416, 99‒103.CrossRefPubMedGoogle Scholar
  10. 10.
    Cai Y., Xu Z., Nagarajan L., Brandt S.J. 2008. Single-stranded DNA-binding proteins regulate the abundance and function of the LIM-homeodomain transcription factor LHX2 in pituitary cells. Biochem. Biophys. Res. Commun. 373, 303‒308.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Prasad K.V., Ao Z., Yoon Y., Wu M.X., Rizk M., Jacquot S., Schlossman S.F. 1997. CD27, a member of the tumor necrosis factor receptor family, induces apoptosis and binds to Siva, a proapoptotic protein. Proc. Natl. Acad. Sci. U. S. A. 94, 6346‒6351.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Spinicelli S., Nocentini G., Ronchetti S., Krausz L.T., Bianchini R., Riccardi C. 2002. GITR interacts with the pro-apoptotic protein Siva and induces apoptosis. Cell. Death Differ. 9, 1382‒1384.CrossRefPubMedGoogle Scholar
  13. 13.
    Du W., Jiang P., Li N., Mei Y., Wang X., Wen L., Yang X., Wu M. 2009. Suppression of p53 activity by Siva1. Cell. Death Differ. 16, 1493‒1504.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Gudi R., Barkinge J., Hawkins S., Prabhakar B., Kanteti P. 2009. Siva-1 promotes K-48 polyubiquitination of TRAF2 and inhibits TCR-mediated activation of NF-kappaB. J. Environ. Pathol. Toxicol. Oncol. 28, 25‒38.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Fan X., Yang Q., Wang Y., Zhang Y., Wang J., Yuan J., Li Y., Wang Y., Deng Y., Yuan W., Mo X., Wan Y., Ocorr K., Yang X., Wu X. 2011. Cloning and characterization of the cardiac-specific Lrrc10 promoter. BMB Rep. 44, 123‒128.CrossRefPubMedGoogle Scholar
  16. 16.
    Johnson J.M., Castle J., Garrett-Engele P., Kan Z., Loerch P.M., Armour C.D., Santos R., Schadt E.E., Stoughton R., Shoemaker D.D. 2003. Genome-wide survey of human alternative pre-mRNA splicing with exon junction microarrays. Science. 302, 2141‒2144.CrossRefPubMedGoogle Scholar
  17. 17.
    Blencowe B.J. 2006. Alternative splicing: New insights from global analyses. Cell. 126, 37‒47.CrossRefPubMedGoogle Scholar
  18. 18.
    Albert M.L., Darnell R.B. 2004. Paraneoplastic neurological degenerations: keys to tumour immunity. Nat. Rev. Cancer. 4, 36‒44.CrossRefPubMedGoogle Scholar
  19. 19.
    Bachinski L.L., Baggerly K.A., Neubauer V.L., Nixon T.J., Raheem O., Sirito M., Unruh A.K., Zhang J., Nagarajan L., Timchenko L.T., Bassez G., Eymard B., Gamez J., Ashizawa T., Mendell J.R., et al. 2014. Most expression and splicing changes in myotonic dystrophy type 1 and type 2 skeletal muscle are shared with other muscular dystrophies. Neuromuscul. Disord. 24, 227‒240.CrossRefPubMedGoogle Scholar
  20. 20.
    Wang X., Zha M., Zhao X., Jiang P., Du W., Tam A.Y., Mei Y., Wu M. 2013. Siva1 inhibits p53 function by acting as an ARF E3 ubiquitin ligase. Nat. Commun. 4, 1551.CrossRefPubMedGoogle Scholar
  21. 21.
    Resch U., Schichl Y.M., Winsauer G., Gudi R., Prasad K., de Martin R. 2009. Siva1 is a XIAP-interacting protein that balances NFkappaB and JNK signalling to promote apoptosis. J. Cell. Sci. 122, 2651‒2661.CrossRefPubMedGoogle Scholar

Copyright information

© Pleiades Publishing, Inc. 2018

Authors and Affiliations

  1. 1.The Center for Heart Development, State Key Lab Developmental Biology of Freshwater Fish, The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, Key Laboratory of Physical Fitness and Exercise, Rehabilitation of Hunan Province, Key Lab of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal UniversityChangshaChina
  2. 2.Birth Health and Genetics Lab, Parenthood Research Institute of Hunan ProvinceChangshaChina
  3. 3.Deptment of Geriatrics, Xiangya Hospital, Central South UniversityChangshaChina
  4. 4.Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical SciencesGuangzhouChina

Personalised recommendations