Epigenetic Silencing of Spermatocyte-Specific and Neuronal Genes by SUMO Modification of the Transcription Factor Sp3

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SUMO modification of transcription factors is linked to repression of transcription. The physiological significance of SUMO attachment to a particular transcriptional regulator, however, is largely unknown. We have employed the ubiquitously expressed murine transcription factor Sp3 to analyze the role of SUMOylation in vivo. We generated mice and mouse embryonic fibroblasts (MEFs) carrying a subtle point mutation in the SUMO attachment sequence of Sp3 (IKEE₅₅₃D mutation). The E₅₅₃D mutation impedes SUMOylation of Sp3 at K₅₅₁ in vivo, without affecting Sp3 protein levels. Expression profiling revealed that spermatocyte-specific genes, such as Dmc1 and Dnahc8, and neuronal genes, including Paqr6, Rims3, and Robo3, are de-repressed in non-testicular and extra-neuronal mouse tissues and in mouse embryonic fibroblasts expressing the SUMOylation-deficient Sp3E₅₅₃D mutant protein. Chromatin immunoprecipitation experiments show that transcriptional de-repression of these genes is accompanied by the loss of repressive heterochromatic marks such as H3K9 and H4K20 tri-methylation and impaired recruitment of repressive chromatin-modifying enzymes. Finally, analysis of the DNA methylation state of the Dmc1, Paqr6, and Rims3 promoters by bisulfite sequencing revealed that these genes are highly methylated in Sp3wt MEFs but are unmethylated in Sp3E₅₅₃D MEFs linking SUMOylation of Sp3 to tissue-specific CpG methylation. Our results establish SUMO conjugation to Sp3 as a molecular beacon for the assembly of repression machineries to maintain tissue-specific transcriptional gene silencing.

Vyšlo v časopise: Epigenetic Silencing of Spermatocyte-Specific and Neuronal Genes by SUMO Modification of the Transcription Factor Sp3. PLoS Genet 6(11): e32767. doi:10.1371/journal.pgen.1001203
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1001203

Souhrn

Zdroje

1. Geiss-FriedlanderR

MelchiorF

2007 Concepts in sumoylation: a decade on. Nat Rev Mol Cell Biol 8 947 956

2. SeelerJS

DejeanA

2003 Nuclear and unclear functions of SUMO. Nat Rev Mol Cell Biol 4 690 699

3. Garcia-DominguezM

ReyesJC

2009 SUMO association with repressor complexes, emerging routes for transcriptional control. Biochim Biophys Acta 1789 451 459

4. GillG

2005 Something about SUMO inhibits transcription. Curr Opin Genet Dev 15 536 541

5. HayRT

2005 SUMO: a history of modification. Mol Cell 18 1 12

6. RossS

BestJL

ZonLI

GillG

2002 SUMO-1 modification represses Sp3 transcriptional activation and modulates its subnuclear localization. Mol Cell 10 831 842

7. SapetschnigA

RischitorG

BraunH

DollA

SchergautM

2002 Transcription factor Sp3 is silenced through SUMO modification by PIAS1. EMBO J 21 5206 5215

8. ValinA

GillG

2007 Regulation of the dual-function transcription factor Sp3 by SUMO. Biochem Soc Trans 35 1393 1396

9. BouwmanP

PhilipsenS

2002 Regulation of the activity of Sp1-related transcription factors. Mol Cell Endocrinol 195 27 38

10. PhilipsenS

SuskeG

1999 A tale of three fingers: the family of mammalian Sp/XKLF transcription factors. Nucleic Acids Res 27 2991 3000

11. SuskeG

1999 The Sp-family of transcription factors. Gene 238 291 300

12. SuskeG

BrufordE

PhilipsenS

2005 Mammalian SP/KLF transcription factors: bring in the family. Genomics 85 551 556

13. DennigJ

BeatoM

SuskeG

1996 An inhibitor domain in Sp3 regulates its glutamine-rich activation domains. EMBO J 15 5659 5667

14. SapetschnigA

KochF

RischitorG

MennengaT

SuskeG

2004 Complexity of translationally controlled transcription factor Sp3 isoform expression. J Biol Chem 279 42095 42105

15. StielowB

SapetschnigA

KrügerI

KunertN

BrehmA

2008 Identification of SUMO-dependent chromatin-associated transcriptional repression components by a genome-wide RNA interference screen. Mol Cell 29 742 754

16. StielowB

SapetschnigA

WinkC

KrügerI

SuskeG

2008 SUMO-modified Sp3 represses transcription by provoking local heterochromatic gene silencing. EMBO Rep 9 899 906

17. SakaiK

MiyazakiJ

1997 A transgenic mouse line that retains Cre recombinase activity in mature oocytes irrespective of the cre transgene transmission. Biochem Biophys Res Commun 237 318 324

18. SamantSA

OgunkuaO

HuiL

FossellaJ

PilderSH

2002 The T complex distorter 2 candidate gene, Dnahc8, encodes at least two testis-specific axonemal dynein heavy chains that differ extensively at their amino and carboxyl termini. Dev Biol 250 24 43

19. YoshidaK

KondohG

MatsudaY

HabuT

NishimuneY

1998 The mouse RecA-like gene Dmc1 is required for homologous chromosome synapsis during meiosis. Mol Cell 1 707 718

20. CamurriL

MambetisaevaE

SundaresanV

2004 Rig-1 a new member of Robo family genes exhibits distinct pattern of expression during mouse development. Gene Expr Patterns 4 99 103

21. LiangF

ZhangB

TangJ

GuoJ

LiW

2007 RIM3gamma is a postsynaptic protein in the rat central nervous system. J Comp Neurol 503 501 510

22. TangYT

HuT

ArterburnM

BoyleB

BrightJM

2005 PAQR proteins: a novel membrane receptor family defined by an ancient 7-transmembrane pass motif. J Mol Evol 61 372 380

23. FuX

KampsMP

1997 E2a-Pbx1 induces aberrant expression of tissue-specific and developmentally regulated genes when expressed in NIH 3T3 fibroblasts. Mol Cell Biol 17 1503 1512

24. BouwmanP

GöllnerH

ElsässerHP

EckhoffG

KarisA

2000 Transcription factor Sp3 is essential for post-natal survival and late tooth development. EMBO J 19 655 661

25. PohlersM

TrussM

FredeU

ScholzA

StrehleM

2005 A role for E2F6 in the restriction of male-germ-cell-specific gene expression. Curr Biol 15 1051 1057

26. StorreJ

SchaferA

ReichertN

BarberoJL

HauserS

2005 Silencing of the meiotic genes SMC1beta and STAG3 in somatic cells by E2F6. J Biol Chem 280 41380 41386

27. KehoeSM

OkaM

HankowskiKE

ReichertN

GarciaS

2008 A conserved E2F6-binding element in murine meiosis-specific gene promoters. Biol Reprod 79 921 930

28. GuentherMG

LevineSS

BoyerLA

JaenischR

YoungRA

2007 A chromatin landmark and transcription initiation at most promoters in human cells. Cell 130 77 88

29. BilodeauS

KageyMH

FramptonGM

RahlPB

YoungRA

2009 SetDB1 contributes to repression of genes encoding developmental regulators and maintenance of ES cell state. Genes Dev 23 2484 2489

30. LunyakVV

BurgessR

PrefontaineGG

NelsonC

SzeSH

2002 Corepressor-dependent silencing of chromosomal regions encoding neuronal genes. Science 298 1747 1752

31. OuyangJ

ShiY

ValinA

XuanY

GillG

2009 Direct binding of CoREST1 to SUMO-2/3 contributes to gene-specific repression by the LSD1/CoREST1/HDAC complex. Mol Cell 34 145 154

32. LiH

RauchT

ChenZX

SzaboPE

RiggsAD

2006 The histone methyltransferase SETDB1 and the DNA methyltransferase DNMT3A interact directly and localize to promoters silenced in cancer cells. J Biol Chem 281 19489 19500

33. SarrafSA

StanchevaI

2004 Methyl-CpG binding protein MBD1 couples histone H3 methylation at lysine 9 by SETDB1 to DNA replication and chromatin assembly. Mol Cell 15 595 605

34. IvanovAV

PengH

YurchenkoV

YapKL

NegorevDG

2007 PHD Domain-Mediated E3 Ligase Activity Directs Intramolecular Sumoylation of an Adjacent Bromodomain Required for Gene Silencing. Mol Cell 28 823 837

35. MaW

HorvathGC

KistlerMK

KistlerWS

2008 Expression patterns of SP1 and SP3 during mouse spermatogenesis: SP1 down-regulation correlates with two successive promoter changes and translationally compromised transcripts. Biol Reprod 79 289 300

36. GöllnerH

DaniC

PhillipsB

PhilipsenS

SuskeG

2001 Impaired ossification in mice lacking the transcription factor Sp3. Mech Dev 106 77 83

37. Van LooPF

BouwmanP

LingKW

MiddendorpS

SuskeG

2003 Impaired hematopoiesis in mice lacking the transcription factor Sp3. Blood 102 858 866