Testin

Protein-coding gene in humans

Not to be confused with testin, the unrelated rat pseudoenzyme.

TES

| Available structures

PDB| Ortholog search: PDBe RCSB
| List of PDB id codes

2IYB, 2XQN

Identifiers
Aliases| TES, TESS, TESS-2, testin LIM domain protein
External IDs| OMIM: 606085; MGI: 105081; HomoloGene: 41051; GeneCards: TES; OMA:TES - orthologs
| Gene location (Human)

Chr.| Chromosome 7 (human)[1]

Band| 7q31.2| Start| 116,210,506 bp[1]
End| 116,258,783 bp[1]
| Gene location (Mouse)

Chr.| Chromosome 6 (mouse)[2]

Band| 6 A2|6 7.67 cM| Start| 17,065,148 bp[2]
End| 17,105,827 bp[2]
| RNA expression pattern

Bgee| | Human| Mouse (ortholog)

|

| Top expressed in

* tail of epididymis

* seminal vesicula

* saphenous vein

* gastric mucosa

* superficial temporal artery

* muscle layer of sigmoid colon

* Descending thoracic aorta

* nipple

* myometrium

* smooth muscle tissue

| Top expressed in

* retinal pigment epithelium

* pyloric antrum

* left colon

* spermatid

* seminiferous tubule

* genital tubercle

* granulocyte

* endothelial cell of lymphatic vessel

* medullary collecting duct

* gastric mucosa

More reference expression data
BioGPS| | n/a

| Gene ontology

Molecular function|

* zinc ion binding
* protein binding
* metal ion binding
* RNA binding
* cadherin binding

Cellular component|

* cell junction
* plasma membrane
* nucleus
* focal adhesion
* cytoplasm
* cytosol
* protein-containing complex

Biological process|

* regulation of cell population proliferation
* negative regulation of cell population proliferation

Sources:Amigo / QuickGO
| Orthologs

Species| Human| Mouse
Entrez| |

26136

21753

Ensembl| |

ENSG00000135269

ENSMUSG00000029552

UniProt| |

Q9UGI8

P47226

RefSeq (mRNA)| |

NM_015641
NM_152829

NM_011570
NM_207176

RefSeq (protein)| |

NP_056456
NP_690042

n/a

Location (UCSC)| Chr 7: 116.21 – 116.26 Mb| Chr 6: 17.07 – 17.11 Mb
PubMed search| [3]| [4]
Wikidata
| View/Edit Human| View/Edit Mouse

|

Testin (also known as TESS) is a protein that in humans is encoded by the TES gene located on chromosome 7.[5] TES is a 47 kDa protein composed of 421 amino acids found at focal adhesions and is thought to have a role in regulation of cell motility.[6] In addition to this, TES functions as a tumour suppressor.[7] The TES gene is located within a fragile region of chromosome 7, and the promoter elements of the TES gene have been shown to be susceptible to methylation – this prevents the expression of the TES protein. TES came to greater prominence towards the end of 2007 as a potential mechanism for its tumour suppressor function was published.

## Domain organisation

[edit]

Tes is composed of the following domains:

Domain Name  | Boundaries  | Domain type

|
|

Cysteine rich domain  | 1–90  | No Homology
PET domain  | 90–200  | PET domain – no structure
Linker .  | 201–233  | no domain
LIM1  | 234–300  | LIM domain
LIM2  | 300–365  | LIM domain
LIM3  | 366–421  | LIM domain

The structures of the Cysteine rich domain and the PET domain are not known. LIM domains, however, are known as modulators of protein interactions.[8] LIM domain consist of 2 zinc fingers separated by 2 hydrophobic amino acids (generally a phenylalanine and then a leucine).

## Binding partners

[edit]

TES does not appear to be an enzyme; rather it is a protein that mediates/regulates cellular functions via protein–protein interactions. Pull down experiments[9] reveal that TES has putative interactions mediated by the indicated domain:

Partner  | Domain  | ref  | Method

|
|
|

mENA/VASP  | LIM3  | [6][9][10] | Yeast two Hybrid, Pull-down assay, Structure, ITC
Arp7a  | ???  | [6] | Yeast two Hybrid
Zyxin  | LIM1  | [6][9] | Yeast two Hybrid, Pull-down assay
Actin  | PET?  | [9] | Pull-down assay
α-Actinin  | PET?  | [9] | Pull-down assay
Paxillin  | PET?  | [9] | Pull-down assay

Garvalov et al. showed that the interaction between TES & zyxin were direct, using recombinant proteins expressed in E. coli.[9]

Some of the potential binding partners (Zyxin, mENA) can be found in focal adhesion complexes; the range of binding partners indicates a potential role for TES in-between 'privileged' Actin polymerisation and focal adhesion contacts to the extracellular matrix. This tallies with the observation that GFP-tagged TES can be seen at focal adhesions.

## TES as a tumour suppressor

[edit]

In December 2007, Boeda, Briggs et al.[10] showed that the third LIM domain of TES displaces Mena from its usual subcellular positions (focal adhesions or the cell leading edge). The ENA/VASP protein family (of which Mena is a member) are anchored to specific proteins within the cell by a peptide motif consisting of a phenylalanine residue, followed by four proline residues – known as a FPPPP motif. It is the EVH1 domains of VASP/EVL proteins that directly contact the FPPPP motif. The precise architecture of TES:MENA binding was revealed by X-ray crystallography, and showed that the 3rd LIM domain of TES covered up the FPPPP binding site within Menas EVH1 domain. Isothermal titration calorimetry showed that TES has a greater affinity for Mena than its canonical FPPPP ligand, as presented in the focal adhesion protein zyxin. Using microscopy it was shown that either over-expression of GFP-tagged TES, or just the tagged third LIM domain displaced Mena from focal adhesions and reduced mean cell velocity.

These finding were significant given that Mena is often over-expressed in cancer cells, and is thought to be partly responsible for cancer cell motility, and therefore a factor in cancer metastasis. TES is conversely often not produced in cancer cells. It is possible that a drug designed to mimic TES's interaction with Mena could be used to prevent metastasis and thus development of secondary tumours in cancer patients. The work was widely reported in the British press (the work was carried out by Cancer Research UK),[11][12][13] and also in the international press.[14][15]

## Conformational change

[edit]

Based on the observations that:[citation needed]

* Mammalian cell derived TES binding Zyxin
* E. coli-produced recombinant TES (rTES) does not bind Zyxin
* An rTES construct composed of residues 201–421 (i.e., the linker and all 3 LIM domains) does bind Zyxin
* The above rTES construct binds an N-terminal rTES construct, consisting of the cysteine rich and PET domains – IE, the two-halves of TES interact with each other.

Garvalov et al. propose that TES exists in two conformational states: A 'closed' state where the N & C halves of TES interact, obscuring the Zyxin binding site in LIM1, and an 'open' state where the Zyxin binding site is accessible and the two halves no-longer interact in the same fashion, if at all. The regulatory mechanism switching between the two states is not presently fully understood.

## Phenotype

[edit]

In RNAi experiments, cells that had impaired TES expression showed an inability to correctly organise their focal adhesions and actin stress fibres.

In gene knockout experiments, transgenic mice lacking both copies of the TES gene displayed an increased susceptibility to tumour formation when challenged with a carcinogen. Mice retaining the TES gene were less susceptible: thus, TES is a tumour suppressor gene.

## References

[edit]

1. ^ a b c GRCh38: Ensembl release 89: ENSG00000135269 – Ensembl, May 2017
2. ^ a b c GRCm38: Ensembl release 89: ENSMUSG00000029552 – Ensembl, May 2017
3. ^ "Human PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
4. ^ "Mouse PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
5. ^ Tatarelli C, Linnenbach A, Mimori K, Croce CM (August 2000). "Characterization of the human TESTIN gene localized in the FRA7G region at 7q31.2". Genomics. 68 (1): 1–12\. doi:10.1006/geno.2000.6272. PMID 10950921.
6. ^ a b c d Coutts AS, MacKenzie E, Griffith E, Black DM (March 2003). "TES is a novel focal adhesion protein with a role in cell spreading". J. Cell Sci. 116 (Pt 5): 897–906\. doi:10.1242/jcs.00278. PMID 12571287. S2CID 26602119.
7. ^ Drusco A, Zanesi N, Roldo C, Trapasso F, Farber JL, Fong LY, Croce CM (August 2005). "Knockout mice reveal a tumor suppressor function for Testin". Proc. Natl. Acad. Sci. U.S.A. 102 (31): 10947–51\. Bibcode:2005PNAS..10210947D. doi:10.1073/pnas.0504934102. PMC 1182460. PMID 16033868.
8. ^ Dawid IB, Breen JJ, Toyama R (April 1998). "LIM domains: multiple roles as adapters and functional modifiers in protein interactions". Trends Genet. 14 (4): 156–62\. doi:10.1016/S0168-9525(98)01424-3. PMID 9594664.
9. ^ a b c d e f g Garvalov BK, Higgins TE, Sutherland JD, Zettl M, Scaplehorn N, Köcher T, Piddini E, Griffiths G, Way M (April 2003). "The conformational state of Tes regulates its zyxin-dependent recruitment to focal adhesions". J. Cell Biol. 161 (1): 33–9\. doi:10.1083/jcb.200211015. PMC 2172870. PMID 12695497.
10. ^ a b Boëda B, Briggs DC, Higgins T, Garvalov BK, Fadden AJ, McDonald NQ, Way M (December 2007). "Tes, a specific Mena interacting partner, breaks the rules for EVH1 binding". Mol. Cell. 28 (6): 1071–82\. doi:10.1016/j.molcel.2007.10.033. PMID 18158903.
11. ^ "Drug target to stop cancer spread". BBC News. 6 October 2010. Retrieved 6 October 2010.
12. ^ Jha A (28 December 2007). "New light shed on how cancers spread". Science. The Guardian. Retrieved 6 October 2010.
13. ^ Fletcher V (28 December 2007). "The Cancer 'Life Saver'". UK News. Express.co.uk. Retrieved 6 October 2010.
14. ^ "Cancer's spreading mechanism found". Science. The Sydney Morning Herald. 29 December 2007. Retrieved 6 October 2010.
15. ^ "Clue found to checking cancer spread". The Times of India. 29 December 2007. Archived from the original on 19 October 2012. Retrieved 6 October 2010.