Mutation Analysis of Large Tumor Suppressor Genes LATS1 and LATS2 Supports a Tumor Suppressor Role in Human Cancer

Protein Cell 2015, 6(1):6–11 DOI 10.1007/s13238-014-0122-4 Protein & Cell

COMMUNICATION Mutation analysis of large tumor suppressor genes LATS1 and LATS2 supports a tumor suppressor role in human cancer

Tian Yu1, John Bachman2, Zhi-Chun Lai1,2,3&

1 Intercollege Graduate Degree Program in Molecular, Cellular and Integrative Biosciences, The Pennsylvania State University, University Park, PA 16802, USA 2 Department of Biology, The Pennsylvania State University, University Park, PA 16802, USA 3 Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA &

Cell Correspondence: [email protected] (Z.-C. Lai)

& Received October 9, 2014 Accepted November 13, 2014

ABSTRACT encodes a Ser/Thr protein kinase and somatic mutations in human LATS1 and LATS2 have been identified in primary Protein In recent years, human cancer genome projects provide tumors (e.g. Murakami et al., 2011; Visser and Yang, 2010). unprecedented opportunities for the discovery of cancer To systematically evaluate how LATS1/2 genes play a genes and signaling pathways that contribute to tumor critical role in human cancer, a mutation analysis has been development. While numerous gene mutations can be carried out. In the Catalogue of Somatic Mutation in Cancer identified from each cancer genome, what these muta- (COSMIC) database, 101 non-synonymous LATS1 somatic tions mean for cancer is a challenging question to mutations have been identified from 9183 unique human address, especially for those from less understood tumor samples (Fig. 1A). Similarly, there are 80 LATS2 putative new cancer genes. As a powerful approach, in non-synonymous mutations out of 9516 samples (Fig. 1B). silico bioinformatics analysis could efficiently sort out Therefore, an overall mutation rate is 1.10% for LATS1 and mutations that are predicted to damage gene function. 0.84% for LATS2. In the cBioPortal database, LATS1 and Such an analysis of human large tumor suppressor LATS2 exhibited similar overall mutation rates, 1.83% (135/ genes, LATS1 and LATS2, has been carried out and the 7390) and 1.50% (111/7390), respectively. The top three results support a role of hLATS1//2 as negative growth highest mutation rates with relative larger total sample regulators and tumor suppressors. size for LATS1 occurred in stomach adenocarcinoma (5.91%, n = 220), uterine corpus endometrial carcinoma (4%, n = KEYWORDS LATS1 & LATS2, hippo signaling, cancer 248), and bladder urothelial carcinoma (3.1%, n = 130). genome, human cancer Meanwhile, the highest LATS2 non-synonymous mutation rate occurred in uterine corpus endometrial carcinoma (5.2%, n = 248), stomach adenocarcinoma (4.1%, n = 220), and lung adenocarcinoma (3.9%, n = 229) (Table S1). INTRODUCTION To determine the mutation distribution across different Hippo signaling plays a crucial role in animal development domains, analysis through Fisher’s exact test shows that and tumorigenesis (Harvey et al., 2013; Yu and Guan, both the kinase domain (P = 0.01075) and proline-rich (P = 2013). As a key regulator of this growth-inhibitory pathway, 0.0312) of LATS1 displayed the highest mutation frequency the large tumor suppressor (Lats)/warts (wts) gene among all the LATS1 domains. The proline-rich domain had 7 mutations in a 31-amino acid (aa) region (2.2 mutations/10 aa), and the kinase domain had 43 mutations in a 306-aa region (1.41 mutations/10 aa) (Fig. 1A). In LATS2, the kinase Electronic supplementary material The online version of this − domain (P = 5.66 × 10 5) and insertion domain (P = 0.03121) article (doi:10.1007/s13238-014-0122-4) contains supplementary had the highest mutation frequency. LATS2 kinase domain material, which is available to authorized users.

1130 aa

/H R1125C

‡ 2

C1083Y

*1089Y 2

Q1116E 1125 1088 aa Q1079E

G1106A

Omi/HtrA2 V1086A D1078Y C

D1086Y 1111 1130

E1067A 1 /Q R1054*

R1082K 1083

D MST1/2

D1048N

T T1041I/I/P

1086 MST1/2

P D

1079 R1043L 0

T E1039K

1048 HM V1057A D1043N 1041

N1038H E1016K

1001 ‡

F S1023C

D998G

P1028A/T 1039

F1015L F

P996L

R1020T R 1001

/fs F1010L

R983L

R995C/L /H 1020

F978L 2 2 R

* K1005N/

F F972L 1015

D994N 983

995 H970N L967M

N994D

R990Q

C953G/F R

P973T 958

Q959H A944V

F950L

VIII-XI CTD

G909R

VIII-XI CTD L914fs

L903I

E920fs 936

G /N

I902L ‡: found in dbSNP

G Auto

W896* 909 E

T913I T Q903* Auto MST1/2

920 T876N

L894I 913

AS A881V

A899fs E S872L

909 MST1/2 883 P T

P882H 876 978 AS AS

A861V

K863E S

Q863E D871Y P

872

G851E D852N

Y862C

R854K Insert

R849L L841F

T851I

L844M

‡ CHK1

W842L

854 P838S R838G

R827T S P R

846

835

R832G

2+ D837H R

H820R

D F810S 837

N D R817G

G823V

/P /P R806* 2

Mg G803C D

827

A810S D

800 817 fs: frame -shift

A805V 833

D800Y Mg

N A

E802K A773T AMOTL2 S803T 810

R790Q 790 R

R E

L793Q 795 809

802 R769W M790T

F770L A

806

773 M782I

R E765D

G787V/A ATP E

765 R767L Y786H 769 R759W

N762S N ATP V729D

762 A748T Cell E

A758S

753 E726K

N /* /L R744Q E722*

N725S ‡ 4 725 K

R737* 734 716

KIBRA &

5 L699V AR K702M

V719I V

719 L693M

697 R697G

M704V V682L

H691fs

‡

/fs E689* R694C G675W

697 A678S V RR 667 788

682

Q678H K665R

694 T673I LIMK1 LIMK1

R MOB MOB E

M669I

689 R645L 682

E652* 644 690 974 681 727 660

R657C RR

Q663L E

Q643E 657

636 799 652 R Counts of methods to show an alteration as damaging: M643I

K662fs RR L625V 660 655 755

*: nonsense mutation nonsense *: 645

657 F641L

V650A R623W

‡

AJUBA AJUBA NTR Kinase domain-VII Insert R

S596R E644G NTR Kinase Domain I

623

K R593C

K607N CDC2 CDC2 620 629 644 N620K

/fs E591*

S P577L Protein

P S E606G

I615T 613 R581C S

598

592 D569G

E594K

G566C

K595N

D564Y

P579S

A561T

E574*

P551L

R558H /L /L

‡

P568L A546V

E550D G554E

N551S G539D

S528L

P533fs PPPY

YAP NEDD4 ITCH

Y531H

A549S

R525C P531fs

S ‡

4 P

G535E 528

W519C P516L

Y506F

F532fs

P506R/L ZYXIN PPPY

G498V

A483T R502C

A497T

P493S

H475Y

N471S

P452H P475L

NUAK1/2 P468S

G448W PAPA

P454L CHK1/2

N463S S P

464 P445L

S P

446

S444P

A428V S438F R415W

M419I

P434R CHK1/2 P414L

H417D

H393fs S

408 A392fs

R391H

S394Y

P377S 394 675

G393A

S S366F

380 P375S

S387F G363S

‡

AJUBA

PPPY

H359L T367I YAP NEDD4 ITCH PPPY 376 396 Q345*

M310V

S336G AUROR-A CHK1/2 PKA

S G324V P ‡

336 S308F

H317Y

S321T

P305L /L P292fs

A309V

S / P301H

R287*

G293D

I288M

V284I

Y277C

S278C

R271H

P266fs

P W268*

P263S 277 278 P258S

T255N/A LCD2 NTR Kinase domain-VII P250S

2 P T

255 /G R252I

G218V

/fs

T P237Q

2 11 ‡

V234L P

246 R233S

G231*

P208L

P210S I220V

E192K

Y200S

P190S/R Q188R P193fs

‡ Y183C

PKA

W178*

R174C

S179L

CHK1/2 S T168M

A161V 172

P158S

P166S

/M I131V

I149T ‡ 135 353

E130K G113E

L109S

Q114R A120G

E100*

A110T

Q105P

R96Q/L

S91L

G92S

‡

R82Q

/* 84

4 2 L L

85 I80fs

I81M S

L78fs P I

I 83

81 L77fs

78 R63Q

P72L A

NF2 LIMK1 NF2 80 E60G NF2

77 Q74R A T59A

77 77

76 ‡

S45Y 88 71

72 89 D56Y

H52Y AUROR-A

45 G40E

E36D UBALCD1 P-rich

A47S A47S

G36W S33L

‡ SL

R28Q

R R35W

LCD1 UBALCD1 LCD2 28 2 R18* /Q R16L V25F R 16 33 A. LATS1 A. B. LATS2 B. had 44 mutations in a 306-aa region (1.48 mutation/10 aa) occurred to enrich mutations in functionally signiﬁcant and LATS2 insertion domain had 9 in 45 aa (2 mutation/10 regions such as the kinase domain to facilitate aa) (Fig. 1B). These data support that selections have tumorigenesis.

Figure 1. Human cancer mutations in LATS1 (A) and LATS2 s cell line (Visser and Yang, 2010). Lats1 LCD1 knockout (B) are mapped to their corresponding open reading mice were born with a low birth rate, from which the frames. Human LATS1 and LATS2 mutation data was collected mouse embryonic ﬁbroblasts displayed chromosomal from Catalogue of Somatic Mutations in Cancer (COSMIC) (top instability and tumorigenesis (Yabuta et al., 2011). Within portions) and cBioPortal (bottom portions) databases. (A LCD1, a short segment called Conserved N-terminal Motif complete list of all mutations can be found in Table S1). All (CNM) (aa 72–89 for LATS1 and 71–88 for LATS2) is non-synonymous mutations are analyzed, using Uniprot iden- important for membrane recruitment and activation of tiﬁer O95835 for LATS1 and Q9NRM7 for LATS2. Synonymous LATS1/2 by Merlin/NF2. The alterations of three highly mutations were not included in this analysis. To evaluate conserved residues in LATS1/2-A77/76P-I81/80T-L85/84P potential impact of individual mutations on LATS1/2 structure prevented its interaction with Merlin/NF2, membrane and function, the following bioinformatics resources were used: localization and activation (Yin et al., 2013). Therefore, 1) SIFT; 2) PROVEAN; 3) PolyPhen-2; and 4) Mutation LATS1-I81M, LATS1-R82Q, and LATS2-P72L mutations Assessor. A color code was used to distinguish mutations that may fail to interact with Merlin/NF2 and consequently are predicted to be damaging by various numbers of the cannot be activated (Fig. 1). The truncated products of methods described above (zero in dark green, one in light LATS1 such as L78fs, R82*, E100*, W178*, and G231* green, two in orange yellow, three in brown, and four in red). “*” may compete with wild-type LATS proteins for the binding indicates nonsense mutation and “‡” for ones found in dbSNP. partners of LCD1. Interestingly, LATS2-S83 within CNM is “fs” is for frame-shift. Blue bars indicate regions involved in phosphorylated by Aurora-A to regulate the centrosomal protein-protein interactions as indicated. Blue triangles identify localization and mitotic activity of LATS2 (Visser and Yang, phosphorylation sites by corresponding protein kinases. Each Cell 2010). Finally, the conserved residues in LCD1 could be small block square indicates one unique mutation sample for & critical for function. Mutations were found at certain con- LATS1/2 from human cancer. served sites in LCD1 which include LATS1/2-R28/R16, S45/S33, L78/L77, and I81/I80. Moreover, N463S, P468S, H475Y, A483T, P493S, R502C, P506R/L, and W519C in Among all the mutations, nonsense and frame-shift LCD2 for LATS1 are predicted to be damaging. P468S is

Protein mutations clearly disrupt LATS1/2 gene function. Thirteen close to the phosphorylation site LATS1-S464 by NUAK, and 10 unique nonsense mutations were found in LATS1 which promotes LATS1 degradation. Additionally, CDC2 and LATS2, respectively. Moreover, 12 and 7 unique frame- phosphorylates S613 of LATS1. CDC2 forms a complex shift mutations were detected in LATS1 and LATS2, with LATS1 in the centrosome and phosphorylation of respectively (Fig. 1). The pattern of equal distribution of S613 occurs during mitosis (Visser and Yang, 2010). LATS1/2 nonsense or frame-shift mutations is consistent Mutations near S613, such as K607N and I615T, may with the idea that LATS1 and LATS2 are tumor suppressor interfere with phosphorylation at this site. Four lesser genes. The percentage of either nonsense or frame-shift deleterious LATS2 mutations were found in LCD2, which mutations among all the mutations for LATS1 and LATS2, contains the phosphorylation sites S408 and S446 by was 17.43% and 10.69%, respectively. Chk1/2 in response to UV damage (Okada et al., 2011). To predict the functional relevance of other non-synony- Next to LCD2, LATS2-S380 is phosphorylated by Aurora A mous mutations in LATS1/2, we performed analyses of during mitosis, which is critical for Aurora A-LATS-Aurora evolutionary conservation and protein structure through four B axis to regulate mitotic progression (Yabuta et al., 2011). different mutation-assessing methods: SIFT, PROVEAN, Moreover, LATS2 S380 is located within an Ajuba-binding PolyPhen-2, and Mutation Assessor (Ng and Henikoff, 2003; region of LATS2 (aa 376–396), which regulates the spindle Adzhubei et al., 2010; Reva et al., 2011; Choi et al., 2012). apparatus formation (Visser and Yang, 2010). Mutations We found that 73.85% (161/218) of missense mutations from like R391H may affect the interactions and cell cycle LATS1 and 77.98% (124/159) from LATS2 are predicted to control. be damaging by at least one method (Fig. 1). In regards to mutations that were considered severe as determined by all PROLINE-RICH REGION four methods, LATS1 had a percentage of 25.69% (56/218) and LATS2 had 31.44% (50/159). LATS1 has a unique proline-rich region (Fig. 1A). Previous studies detected phosphorylation of T246 and T255 in P-stretch, as well as S336 located downstream (Hornbeck LCD1 AND LCD2 et al., 2012). T255A/N and S336G mutations in LATS1 LATS1/2 proteins share LATS conserved domain 1 (LCD1) would prevent phosphorylation of these residues, and the and LATS conserved domain 2 (LCD2), which are con- R252I/G mutations nearby may affect these phosphoryla- served in all vertebrate LATS1/2 homologues. LCD1 and tion events. Moreover, Y277 and S278 located within the LCD2 are critical for LATS1/2 function and regulation. The LIMK-binding site of LATS1 were also found to be phos- deletions of either LCD1 or LCD2 in mouse Lats2 abol- phorylated (Visser and Yang, 2010). While the functional ished its tumor suppressor activity in immortalized mouse signiﬁcance of these phosphorylations is unknown, Y277C

8 © The Author(s) 2014. This article is published with open access at Springerlink.com and journal.hep.com.cn Mutation analysis of human LATS1 and LATS2 genes COMMUNICATION

and S278C mutations clearly prevent phosphorylation of these conserved catalytic elements, LATS1 V719I/A, these two residues. R744Q/L, A748T, R827T, R837H, L844M and LATS2 G675W, A678S, V682L, L693M, L699V, D800Y, G803C are most likely to disrupt ATP binding and catalysis. LATS1- BINDING WITH YAP AND KIBRA R995C/L/H and LATS2-G909R also change highly con- The interaction between the Proline-Proline-x-Tyrosine served residues. Therefore, these cancer mutants may affect (PPxY) motif and the hydrophobic pocket of WW domain their kinase activity. Moreover, conserved residues in the is critical for the LATS1/2 binding with either their substrate kinase domain of LATS1/2 are expected to be important for YAP or their activator KIBRA. It has been reported that their kinase activity. Mutations occurring in these residues LATS1-Y559F and Y376A abrogated the binding with YAP included LATS1-N762/LATS2-N725, R806/R769, A810/ (Visser and Yang, 2010), while LATS2-Y518A partially A773, R827/R790, D837/D800, R854/R817, and R995/R958 abolish the binding with KIBRA (Xiao et al., 2011). These (Fig. 1). interactions may be affected by the third proline mutation Unlike other AGC family members, LATS1/2 and the P375S in the ﬁrst PPxY motif of LATS1 and the second NDR subfamily members have an insert between the proline mutation P516L in the only PPxY motif of LATS2. kinase subdomain VII and the activation segment (AS) in the subdomain VIII. Basic residues in the insert inhibit the activity of AS likely through an auto-inhibitory mechanism BINDING WITH MOB, AJUBA AND AMOTL2 (Hergovich et al., 2006). Missense mutations in this region The N-terminal regulatory domain (NTR) adjacent to the (e.g. LATS1-D871Y) may have an impact on the kinase kinase domain is required for LATS1/2 activation by MOB, activity (Fig. 1). Cell

AJUBA, and AMOTL2 (Visser and Yang, 2010; Paramasi- & vam et al., 2011; Xiao et al., 2011). Positively charged residues such as LATS1-R657/LATS2-K620, R660/R623, R682/ PHOSPHORYLATION AND ACTIVATION BY MST1/2 R645, R694/R657, and R697/R660 in LATS1/2 NTR are KINASES conserved in the NDR subfamily from yeast to human.

Previous studies have shown that LATS1 mutations (e.g. The phospho-S909/S872 and other residues in AS organize Protein R657A, R660A and R694A) abolish their interaction with the interaction between ATP and substrates of LATS1/2. Previ- negatively charged surface of MOB1A/B and kinase activity ous study has shown that LATS1 mutant S909A/D/E cannot (Hergovich et al., 2006). Therefore, LATS1 mutants (R657C, be activated by the MST2 kinase. Therefore, LATS2-S872L R694C and R697G) and LATS2 mutants (R623W and would abolish the kinase activity, and LATS1-T913I and R645L), are expected to become inactive in human cancer LATS2-T876N, A881V also likely damage catalytic activity. In due to loss of interactions with LATS activators such as the C-terminal of kinase IX and XI subdomains, the LATS2- MOB. The Drosophila Wts-R702 residue is equivalent to G909 and LATS1-R995 are highly conserved among LATS1/2-R694/R657 and critical for Mob-binding, kinase eukaryotic kinases (Endicott et al., 2012). LATS1-R995C/L/H activity, and inhibition of tissue growth in development (Ho and LATS2-G909R would probably damage kinase activity. et al., 2010). LATS2-G909R and C953* has been experimentally shown to be defective for kinase activity and YAP regulation (Yu et al., 2013). THE KINASE DOMAIN In the C-terminal domain (CTD) of AGC family kinase, LATS1 and LATS2 are members of the AGC (named after the conserved NFD (Asn-Phe-Asp) motif interacts with PKA, PKG, and PKC) protein kinase family. Although crystal hydrophobic pocket in the N-terminus of kinase domain to structures of some AGC proteins have been deciphered, facilitate kinase activation (e.g. PKC, Leonard et al., 2011). there is no structure information available for LATS1/2 kinase In LATS1 NFD, N1038H may disrupt the kinase activity. In domains. To estimate mutation-induced structural changes, addition, LATS1-R1020T, P1028A/T, S1023C and LATS2- we performed structure remodeling for LATS1/2 based on P996L, D998G may also affect this activation. LATS1/2 two AGC family proteins, ROCK and PKC. Conserved in activation also requires phosphorylation of T1079/T1041 most eukaryotic protein kinases, the N-terminal catalytic by MST1/2 in hydrophobic motif (HM), which has a con- domain of LATS1/2 interacts with the phosphate donor ATP sensus sequence F-x-x-Y/F-T-Y/F-K/R in the NDR protein through a crucial network, composited by GxGxxGxV loop subfamily (Hergovich et al., 2006). Thus, LATS2-T1041I/P (LATS1: 712–719/ LATS2: 675–682), K734/697, E753/716, mutations clearly abolish phosphorylation at this site to DxKxxN (828–833/791–795) and DFG motif (846–848/809– cause kinase inactivation. LATS1-R1082K, D1086Y and 811) (“x” represents any aa) (Endicott et al., 2012; Hanks LATS2-E1039K, R1043L, D1048N may disrupt kinase and Hunter, 1995). In addition, previous studies also have activity as well. Mutations at conserved sites between veriﬁed that mutants LATS1-K734M/LATS2-K697A/mLats2- LATS1/2 such as LATS1-F1015/LATS2-F978, R1020/R983, K655M are all kinase-dead (Zhao et al., 2007; Wei et al., F1039/F1001, and D1086/D1048 may affect function of the 2007; Visser and Yang, 2010). Mutated within or close to C-terminus.

CONCLUDING REMARKS reproduction in any medium, provided the original author(s) and the source are credited. Genetic analysis of Lats/Wts family genes using Drosophila and mice models has revealed their role as negative growth regulators and tumor suppressors in animal (Visser and REFERENCES Yang, 2010; Harvey et al., 2013; Yu and Guan, 2013). The fact that human LATS1 can functionally replace Wts in Adzhubei IA, Schmidt S, Peshkin L, Ramensky VE, Gerasimova A, Drosophila supports that LATS may function as a tumor Bork P, Kondrashov AS, Sunyaev SR (2010) A method and suppressor in human cells. From human cancer genome server for predicting damaging missense mutations. Nat Methods – projects, an increasing number of mutations in LATS1/2 are 7:248 249 detected. Compared to some well-established cancer genes Choi Y, Sims GE, Murphy S, Miller JR, Chan AP (2012) Predicting such as TP53 and Rb, LATS1 and LATS2 are not frequently the functional effect of amino acid substitutions and indels. PLoS One 7:e46688 mutated. However, our in silico analysis provides supporting Endicott JA, Noble ME, Johnson LN (2012) The structural basis for evidence that LATS1/2 mutations drive human tumor control of eukaryotic protein kinases. Annu Rev Biochem 81:587– development based on the following observations: 1) Cancer 613 mutations in hLATS1/2 do not appear to be random muta- Hanks SK, Hunter T (1995) Protein kinases 6. The eukaryotic protein tions. Damaging mutations have been accumulated more kinase superfamily: kinase (catalytic) domain structure and preferentially in important protein domains such as the classiﬁcation. FASEB J 9:576–596 kinase domain. Majority of the mutations including nonsense Harvey KF, Zhang X, Thomas DM (2013) The Hippo pathway and

Cell and frame-shift mutations clearly disrupt LATS1/2 function; human cancer. Nat Rev Cancer 13:246–257 2) Some mutations occurred in regions and residues & Hergovich A, Stegert MR, Schmitz D, Hemmings BA (2006) NDR important for LATS1/2 activation. Examples include MST1/2 kinases regulate essential cell processes from yeast to humans. phosphorylation sites, MOB-binding domain, and the region Nat Rev Mol Cell Biol 7:253–264 critical for NF2 interaction; 3) In certain cancer types like Ho LL, Wei X, Shimizu T, Lai ZC (2010) Mob as tumor suppressor is stomach adenocarcinoma and uterine corpus endometrial activated at the cell membrane to control tissue growth and organ carcinoma, mutation rates can be reasonably high (5.2%– Protein size in Drosophila. Dev Biol 337:274–283 5.9%). On the other hand, mechanisms other than gene Hornbeck PV, Kornhauser JM, Tkachev S, Zhang B, Skrzypek E, mutation could also be effective to alter gene activity. In this Murray B, Latham V, Sullivan M (2012) PhosphoSitePlus: a regard, LATS1/2 genes are known to be down-regulated by comprehensive resource for investigating the structure and promoter methylation. Mutations in other Hippo pathway function of experimentally determined post-translational modifi- genes such as MST1/2 are also expected to reduce LATS1/2 cations in man and mouse. Nucleic Acids Res 40:D261–D270 function. While the functional significance of some of these Leonard TA, Różycki B, Saidi LF, Hummer G, Hurley JH (2011) mutations have been experimentally tested in vivo (Yu et al., Crystal structure and allosteric activation of protein kinase C βII. 2013), results reported here further support that LATS1/2 act Cell 144:55–66 normally as tumor suppressors and loss of their functions Murakami H, Mizuno T, Taniguchi T, Fujii M, Ishiguro F, Fukui T, contributes to human cancer development. Akatsuka S, Horio Y, Hida T, Kondo Y, Toyokuni S, Osada H, Sekido Y (2011) LATS2 is a tumor suppressor gene of malignant ACKNOWLEDGEMENTS mesothelioma. Cancer Res 71:873–883 Ng PC, Henikoff S (2003) SIFT: predicting amino acid changes that This work was partly supported by the National Science Foundation. affect protein function. Nucleic Acids Res 31:3812–3814 We would like to thank Nicholas Anzalone and Samar Almarzooqi for Okada N, Yabuta N, Suzuki H, Aylon Y, Oren M, Nojima H (2011) A assistance and other members of the Lai laboratory for discussions. novel Chk1/2–Lats2–14-3-3 signaling pathway regulates P-body formation in response to UV damage. J Cell Sci 124:57–67 ABBREVIATIONS Paramasivam M, Sarkeshik A, Yates JR, Fernandes MJ, McCollum CTD, C-terminal domain; HM, hydrophobic motif; LCD1, LATS D (2011) Angiomotin family proteins are novel activators of the conserved domain 1; LCD2, LATS conserved domain 2; NTR, LATS2 kinase tumor suppressor. Mol Biol Cell 22:3725–3733 N-terminal regulatory domain; PPxY, proline-proline-x-tyrosine. Reva B, Antipin Y, Sander C (2011) Predicting the functional impact of protein mutations: application to cancer genomics. Nucleic COMPLIANCE WITH ETHICS GUIDELINES Acids Res 39:e118 Visser S, Yang X (2010) LATS tumor suppressor: a new governor of Tian Yu, John Bachman, and Zhi-Chun Lai declare that they have no cellular homeostasis. Cell Cycle 9:3892–3903 conflict of interest. This article does not contain any studies with Wei X, Shimizu T, Lai ZC (2007) Mob as tumor suppressor is human or animal subjects performed by the any of the authors. activated by Hippo kinase for growth inhibition in Drosophila. EMBO J 26:1772–1781 OPEN ACCESS Xiao L, Chen Y, Ji M, Dong J (2011) KIBRA regulates Hippo This article is distributed under the terms of the Creative Commons signaling activity via interactions with large tumor suppressor Attribution License which permits any use, distribution, and kinases. J Biol Chem 286:7788–7796

10 © The Author(s) 2014. This article is published with open access at Springerlink.com and journal.hep.com.cn Mutation analysis of human LATS1 and LATS2 genes COMMUNICATION

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