Pereira Sap 2008 Ndpk

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Sociedad Argentina de Protozoologa

02-05 de Noviembre de 2008

Tr anosoma cruziTr anosoma cruzi

nucleoside diphosphatenucleoside diphosphate

Claudio A. Pereira

Laboratorio de Biologa Molecular de Trypanosoma cruziInstituto de Investigaciones Medicas A. Lanari

Universidad de Buenos Aires - CONICET


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arginine+ATP phosphoarginine+ ADP

Adenylatekinase

2ADP ATP+AMP

N1TP + N

2DP N

1DP + N

2TP


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easons o s u y p osp o ransp erases: easons o s u y p osp o ransp erases:

. ere are many p osp otrans erase o trypanosomat s a sent

in mammals.

.

nucleotide interconversion

structures such as flagella and nucleus

4. The movement of ATP within a cell is not completely clarified


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Far more than HOUSEKEEPING ENZYMES, other than the

osynt et c processes:

1. Signal transduction by providing GTP to G proteins

activation.

2. Signal transduction by transference of the phosphohistidine

phosphate to different acceptors as part of a

phosphorylation cascade. It was also involved in bacterialwo-componen s gna rans uc on sys ems.

3. Several functions in cell development, proliferation,

differentiation, tumor metastasis, apoptosis and motility

.4. DNA processing: in humans NM23-H2/NDPK B is a

transcription factor with sequence-specific DNA binding

. ,

the DNA targets suggest a possible participation in DNA

repair.


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199

5

TcNDPK1?Nuclear TbNDPK1


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TriTr s TriTr s DATA MINING:

TcNDPK2: DM10-containing long isoform

c : putat ve em c

TcNDPK4: putative atypical long isoform


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proac

proac

firsta

firsta

.cruzi:

.cruzi:

sdesde

NDPNDP


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onon

func

t

func

t

an

ove

an

ove

PK1:

PK1:

TcNTcN


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tsts

nemn

nemn

K

sali

K

sali

TcND

TcND


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......

s

Crysta

istance

NDPK

BZLre

T


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TcNDPK2: What is the role of DM10 motifs?TcNDPK2: What is the role of DM10 motifs?

A novel Trypa nosoma c ruzinucleoside diphosphate kinase

targeted to the flage llum

Mariana R. Mirand a, Len A. Bouvier, Gaspar E. Canep a a nd Cla udio A.

Pereira *

Laborat orio de Biologa Mo lecula r de Trypan osoma c ruzi (LBMTC), Instituto d e

Investiga cione s Md ica s Alfredo Lanari, Universidad de Buenos Aires and C ONICET,

Buenos Aires, Argentina .

DM10: Domains in hypothetical proteins in Drosophila, C. elegansand mammals. Occurs singly in some nucleoside diphosphate kinases.

NDPK2

E-value: 4e-08

DM10-TcNDPK2 6 RYAFKVDYYDPQA--NLTRQYLLLYYTEDATIEMHDLKTK------RVFLKRCAYPS---------LGVQELFIGATVntDi

smart00676 4 VLRFDAYWDDPVAmpGLIRRFKISYYLEDDTIEIFEPDVRnsgilgGTFLRRQRVPKpsppddpefYTASDLNVGTTIN

DM10-TcNDPK2 69 FSRSLKLVDYgDEATRQ 85

smart00676 84 FGRQFRIYDC-DEFTRN 99

NDPK3

E-value: 3e-07

DM10-TcNDPK3 9 RLALYCERQDDISQ--LVRRFVLFFFYEDRSIEMRE-IPKNV-----LYLRRAPFP---------HLKKDDFTLGASLTalignm

itonin

smar pg p gi gg pspp pe

DM10-TcNDPK3 72 NGGIVKITDYaDEVTRVLCEKKS 94

smart00676 84 FGRQFRIYDC-DEFTRNYLESKG 105

NDPK2 vs NDPK3

Expect = 3e-13

Identities = 25/79 (31%), Positives = 45/79 (56%)

TcNDPK2 6 RYAFKVDYYDPQANLTRQYLLLYYTEDATIEMHDLKTKRVFLKRCAYPSLGVQELFIGAT 65

TcNDPK3 9 RLALYCER DDIS LVRRFVLFFFYEDRSIEMREIPKNVLYLRRAPFPHLKKDDFTLGAS 68

DM10

xtractio

TcNDPK2 66 VNIFSRSLKLVDYGDEATR 84

TcNDPK3 69 LTINGGIVKITDYADEVTR 87


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DM10 motifs participate in the TcNDPK localizationDM10 motifs participate in the TcNDPK localization

TcNDPK2 (WT, CE)TcNDPK2 (WT, WC)

PAR2 (WT, CE)GFP (DM10-GFP, CE)

PAR-2 TcNDPK2


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TcNDPK2 regulation mechanismsTcNDPK2 regulation mechanisms

Glycosidases Phosphatases Trans-splice mapping

NO NO ?


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Additional DataAdditional Data

TcNDPK2TcNDPK2

TcNDPK3TcNDPK3


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......Ex lore the interrelations of different hos hotransferases in

the cellValidation of the different TcNDPK isoforms

Addressing the functional aspects of TcNDPK1 in relation

with its nuclease activity, BZL resistance and TcNDPK2 inflagella

Study the stage-specific and other regulation factors


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LAB MEMBERS

..

Leon Bouvier (ADKs)

Gaspar Canepa (Transporters)

CCLabora

Labora

Instit

Instit

ationa co a orations

Martn Vzquez (FCEyN,UBA)

Carolina Carrillo (Instituto Leloir) RKGRO

U

SS

toriode

toriode

TrypTryp

todeItodeI

DIM(DIM(

Esteban Serra (IBR, Rosario)

Antonio Uttaro (IBR, Rosario)

Carlos Aguilar (UNSL, San Luis) TW

ORKW

OOBiolog

Biolog

nosoma

nosoma

vestiga

vestiga

ON

IC

ON

IC

International collaborations

Michael Ginger (Lancaster University)

N

UTUT

aMole

c

aMole

c

cruzi

cruzi

iones

iones

TT--UBA

UBA

SUPPORT

ANPCyT: PICT RED 0300, PICT 33431

ularde

ularde

edicas

edicas

CONICET: PIP RED 5492

UBA: UBACyT X073

Pereira Sap 2008 Ndpk

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