Subcellular Localization of Different Regions of Porcine Six1 Gene and Its Expression Analysis in C2C12 Myoblasts

Overview

Journal Mol Biol Rep

Specialty Molecular Biology

Date 2012 Jul 4

PMID 22752728

Citations 5

Authors

Wangjun Wu

Zhuqing Ren

Chao Chen

Yang Liu

Lin Zhang

Zhe Chao

Bo Zuo

Dequan Xu

Minggang Lei

Yuanzhu Xiong

Affiliations

Soon will be listed here.

Abstract

Six1 protein belongs to the Six homeoproteins family, exposing typical domain structure. Although the functions of Six1 have been drawn much attention, the roles of its individual domains are not completely elucidated. Here, we first detected the expression patterns of myogenin, MyoD, Myf5, and Six1 genes using real-time PCR in differentiating C2C12 cells cultured in differentiation medium for 2 or 6 days. The results showed that Six1 gene had the similar expression pattern with myogenin, MyoD, and Myf5 genes, which suggests that it may affect the myogenic differentiation. In order to evaluate the role of distinct domains of Six1 protein in subcellular localization, we constructed a series of truncated vectors tagged with green fluorescent proteins expressing various regions of porcine Six1 protein for subcellular localization analysis. Fluorescence confocal microscopy analysis showed that the different regions of Six1 protein displayed discrete distributions throughout the nucleus and the cytoplasm. The full-length CDS was exclusively localized in the nucleus and the individual HD domain was preferentially distributed to the nucleus both in C2C12 cells and in PK cells. However, the SD domain was diffusely distributed to the cytoplasm and the nucleus, and the localization of SD domain was biased to cytoplasm in C2C12 cells. Taken together, we conclude that the HD domain is important for the nuclear localization of porcine Six1 protein.

Citing Articles

Cytoplasmatic Localization of Six1 in Male Testis and Spermatogonial Stem Cells.

Qin M, Ma L, Du W, Chen D, Luo G, Liu Z Int J Stem Cells. 2024; 17(3):298-308.

PMID: 38225887 PMC: 11361848. DOI: 10.15283/ijsc23093.

Modeling the transport of nuclear proteins along single skeletal muscle cells.

Taylor-Weiner H, Grigsby C, Ferreira D, Dias J, Stevens M, Ruas J Proc Natl Acad Sci U S A. 2020; 117(6):2978-2986.

PMID: 31988126 PMC: 7022209. DOI: 10.1073/pnas.1919600117.

MicroRNA-203 suppresses proliferation in liver cancer associated with PIK3CA, p38 MAPK, c-Jun, and GSK3 signaling.

Zhang A, Lakshmanan J, Motameni A, Harbrecht B Mol Cell Biochem. 2017; 441(1-2):89-98.

PMID: 28887744 DOI: 10.1007/s11010-017-3176-9.

LncRNA-Six1 Encodes a Micropeptide to Activate in and Is Involved in Cell Proliferation and Muscle Growth.

Cai B, Li Z, Ma M, Wang Z, Han P, Abdalla B Front Physiol. 2017; 8:230.

PMID: 28473774 PMC: 5397475. DOI: 10.3389/fphys.2017.00230.

Overexpression of sineoculis homeobox homolog 1 predicts poor prognosis of hepatocellular carcinoma.

Kong J, Zhou X, Liu S, Jin T, Piao Y, Liu C Int J Clin Exp Pathol. 2014; 7(6):3018-27.

PMID: 25031720 PMC: 4097294.

References

Braun T, Rudnicki M, Arnold H, Jaenisch R . Targeted inactivation of the muscle regulatory gene Myf-5 results in abnormal rib development and perinatal death. Cell. 1992; 71(3):369-82. DOI: 10.1016/0092-8674(92)90507-9. View

Chen J, Goldhamer D . Transcriptional mechanisms regulating MyoD expression in the mouse. Cell Tissue Res. 1999; 296(1):213-9. DOI: 10.1007/s004410051282. View

Rudnicki M, Schnegelsberg P, Stead R, Braun T, Arnold H, Jaenisch R . MyoD or Myf-5 is required for the formation of skeletal muscle. Cell. 1993; 75(7):1351-9. DOI: 10.1016/0092-8674(93)90621-v. View

Rudnicki M, Braun T, Hinuma S, Jaenisch R . Inactivation of MyoD in mice leads to up-regulation of the myogenic HLH gene Myf-5 and results in apparently normal muscle development. Cell. 1992; 71(3):383-90. DOI: 10.1016/0092-8674(92)90508-a. View

Maroto M, Reshef R, Munsterberg A, Koester S, Goulding M, Lassar A . Ectopic Pax-3 activates MyoD and Myf-5 expression in embryonic mesoderm and neural tissue. Cell. 1997; 89(1):139-48. DOI: 10.1016/s0092-8674(00)80190-7. View

Kablar B, Krastel K, Ying C, Tapscott S, Goldhamer D, Rudnicki M . Myogenic determination occurs independently in somites and limb buds. Dev Biol. 1999; 206(2):219-31. DOI: 10.1006/dbio.1998.9126. View

Grifone R, Laclef C, Spitz F, Lopez S, Demignon J, Guidotti J . Six1 and Eya1 expression can reprogram adult muscle from the slow-twitch phenotype into the fast-twitch phenotype. Mol Cell Biol. 2004; 24(14):6253-67. PMC: 434262. DOI: 10.1128/MCB.24.14.6253-6267.2004. View

Ohto H, Kamada S, Tago K, Tominaga S, Ozaki H, Sato S . Cooperation of six and eya in activation of their target genes through nuclear translocation of Eya. Mol Cell Biol. 1999; 19(10):6815-24. PMC: 84678. DOI: 10.1128/MCB.19.10.6815. View

Spitz F, Demignon J, Porteu A, Kahn A, Concordet J, Daegelen D . Expression of myogenin during embryogenesis is controlled by Six/sine oculis homeoproteins through a conserved MEF3 binding site. Proc Natl Acad Sci U S A. 1998; 95(24):14220-5. PMC: 24354. DOI: 10.1073/pnas.95.24.14220. View

10.

Weintraub H . The MyoD family and myogenesis: redundancy, networks, and thresholds. Cell. 1993; 75(7):1241-4. DOI: 10.1016/0092-8674(93)90610-3. View

11.

Niro C, Demignon J, Vincent S, Liu Y, Giordani J, Sgarioto N . Six1 and Six4 gene expression is necessary to activate the fast-type muscle gene program in the mouse primary myotome. Dev Biol. 2009; 338(2):168-82. DOI: 10.1016/j.ydbio.2009.11.031. View

12.

Xu P, Zheng W, Huang L, Maire P, Laclef C, Silvius D . Six1 is required for the early organogenesis of mammalian kidney. Development. 2003; 130(14):3085-94. PMC: 3872112. DOI: 10.1242/dev.00536. View

13.

Seo H, Curtiss J, Mlodzik M, Fjose A . Six class homeobox genes in drosophila belong to three distinct families and are involved in head development. Mech Dev. 1999; 83(1-2):127-39. DOI: 10.1016/s0925-4773(99)00045-3. View

14.

Ozaki H, Nakamura K, Funahashi J, Ikeda K, Yamada G, Tokano H . Six1 controls patterning of the mouse otic vesicle. Development. 2003; 131(3):551-62. DOI: 10.1242/dev.00943. View

15.

Giordani J, Bajard L, Demignon J, Daubas P, Buckingham M, Maire P . Six proteins regulate the activation of Myf5 expression in embryonic mouse limbs. Proc Natl Acad Sci U S A. 2007; 104(27):11310-5. PMC: 2040895. DOI: 10.1073/pnas.0611299104. View

16.

Wu W, Ren Z, Wang Y, Chao Z, Xu D, Xiong Y . Molecular characterization, expression patterns and polymorphism analysis of porcine Six1 gene. Mol Biol Rep. 2010; 38(4):2619-32. DOI: 10.1007/s11033-010-0403-9. View

17.

Bessarab D, Chong S, Srinivas B, Korzh V . Six1a is required for the onset of fast muscle differentiation in zebrafish. Dev Biol. 2008; 323(2):216-28. DOI: 10.1016/j.ydbio.2008.08.015. View

18.

Bhagavati S, Song X, Siddiqui M . RNAi inhibition of Pax3/7 expression leads to markedly decreased expression of muscle determination genes. Mol Cell Biochem. 2007; 302(1-2):257-62. DOI: 10.1007/s11010-007-9444-3. View

19.

Grifone R, Demignon J, Houbron C, Souil E, Niro C, Seller M . Six1 and Six4 homeoproteins are required for Pax3 and Mrf expression during myogenesis in the mouse embryo. Development. 2005; 132(9):2235-49. DOI: 10.1242/dev.01773. View

20.

Kucharczuk K, Love C, Dougherty N, Goldhamer D . Fine-scale transgenic mapping of the MyoD core enhancer: MyoD is regulated by distinct but overlapping mechanisms in myotomal and non-myotomal muscle lineages. Development. 1999; 126(9):1957-65. DOI: 10.1242/dev.126.9.1957. View