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HOG model

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Variable Type Variable name

External EGlu,EGln,EAsp,

metabolite EGly,EAla,EAsn,

EArg,ESer,ECys,

EHis,EThr,ETrp,

EPhe,EVal,EMet,

ELeu,EIle,ETyr,

EPro,ELys

ENH3

Internal IGlu,IGln,IAsp,

metabolite IGly,IAla,IAsn,

IArg,ISer,ICys,

Ihis,IThr,ITrp,

IPhe,IVal,IMet,

ILeu,IIle,ITyr,Ipro



INH3



IAlphaKetoGlutarate





IHomoCytrate





IAlphaASAA





ILys









biosynthesis regulation

mRNA GCN2m

GCN4m

LYS14m

MKS1m

active protein GCN2ap





GCN4ap





LYS14ap





MKS1ap







Translation Initiation complex

mRNA GCN3m

GCD1m

GCD6m

GCD7m

GCD2m

GCD11m

SUI2m

SUI3m

GCN20m

GCN1m

active Protein GCN3ap

GCD1ap

GCD6ap

GCD7ap

GCD2ap

GCD11ap

SUI2ap

SUI3ap

GCN20ap

GCN1ap





Lysine biosynthesis pathway enzymes

mRNA LYS1m

LYS2m

LYS4m

LYS5m

LYS9m

LYS12m

LYS20m

LYS21m

YJL200Cm

active Protein LYS1ap

LYS2ap



LYS4ap

LYS5ap

LYS9ap

LYS12ap

LYS20ap



LYS21ap



YJL200Cap





Permeases

mRNA GAP1m

CAN1m

PUT4m

MUP3m

DIP5m

SAM3m

AGP3m

MUP1m

HIP1m

APL1m

MMP1m

AGP2m

LYP1m

AGP1m

GNP1m

TAT2m

TAT1m

BAP2m

BAP3m

MEP1m

MEP2m

MEP3m



active proteinsGAP1ap

CAN1ap

PUT4ap

MUP3ap

DIP5ap

SAM3ap

AGP3ap

MUP1ap

HIP1ap

APL1ap

MMP1ap

AGP2ap

LYP1ap

AGP1ap

GNP1ap

TAT2ap

TAT1ap

BAP2ap

BAP3ap

MEP1ap

MEP2ap

MEP3ap







mRNA SSYm

GRR1m

GLN3m

GAT1m

active proteinsSSYap

GRR1ap

GLN3ap



GAT1ap







bibliography

1 A. G. Hinnebusch

2 B. Regenberg and L.D. Morten and C. Kielland-Brandt and S. Holmberg

3 H. Forsberg and P.O. Ljungdahl

4 B. Magasanik and C. A. Kaiser

5 J. Hofman-Bang

6 G. Giaever et al.

7 L. A. Urrestarazu, C. W. Borell and J. K. Bhattacharjee

15 Hinnebusch AG, Natarajan K.

16 Cigan AM, Bushman JL, Boal TR, Hinnebusch AG.

10 O. Grundman, H.U. Mosch and H. Braus

11 A. Feller, F. Ramos, A. Pierard and E. Dubois

12 A. Feller and F. Ramos and A. Pierard and E. Dubois

13 J. J. Tate and K. H. Cox and R. Rai and T. G. Cooper

14 F. Ramos and E. Dubois and A. Pierard

Description

amino acids









external ammonia

internal amino acids. Each amino acid has certain permeases

(aa_permeases) detailed below.









internal ammonia



metabolite in lysine biosynthesis pathway.

In general, the functions follow these rules:

If there is flow into it, but no flow out, the metabolite is absent (0).

If there is flow in and out, we set its value to 1.

If there is no flow into it. The metabolite is accumulate (2)









protein kinase

amino acids biosynthetic enzymes transcription regulator

lysine biosynthetic enzymes transcription regulator

negative regulator of CIT2 gene expression needed for alphaKetoglutarate synthesis









subunits of the translation initiation complex eIF3

hesis pathway enzymes









lysine biosynthetic enzymes









*replacing LYS7, see ref 6.C170







permease of Asp,Lys,Gly,Ala,Asn,Arg,Ser,Cys,his,Thr,Trp,Phe,Val,Met,Leu,Ile,Tyr

permease of Arg

permease of Ala,Val,Pro

permease of Met

permease of Asp,Glu,Gln,Asn,Ser

permease of Met

permease of Asp,Glu,Ser

permease of Met

permease of His

permease of Arg

permease of Met

permease of Leu.Thr

permease of Lys

permease of Ala,Gly,Val,Ile,Leu,Phe.Met.Cys,Gln,Asn,Ser,Thr,Tyr

permease of Leu,Met,Cys,Glu,Gln,Asn,Ser,Thr

permease of Ala,Gly,Phe,Trp,Tyr

permease of Val,Trp,Thr,Tyr

permease of Leu,Ile,Val,Ala,Phe,Trp,Met,Cys,Tyr

permease of Leu,Ile,Val,Ala,Phe,Trp,Met,Cys,Tyr,Thr

permease of ammonia

permease of ammonia

permease of ammonia



permease of Asp,Lys,Gly,Ala,Asn,Arg,Ser,Cys,his,Thr,Trp,Phe,Val,Met,Leu,Ile,Tyr

permease of Arg

permease of Ala,Val,Pro

permease of Met

permease of Asp,Glu,Gln,Asn,Ser

permease of Met

permease of Asp,Glu,Ser

permease of Met

permease of His

permease of Arg

permease of Met

permease of Leu.Thr

permease of Lys

permease of Ala,Gly,Val,Ile,Leu,Phe.Met.Cys,Gln,Asn,Ser,Thr,Tyr

permease of Leu,Met,Cys,Glu,Gln,Asn,Ser,Thr

permease of Ala,Gly,Phe,Trp,Tyr

permease of Val,Trp,Thr,Tyr

permease of Leu,Ile,Val,Ala,Phe,Trp,Met,Cys,Tyr

permease of Leu,Ile,Val,Ala,Phe,Trp,Met,Cys,Tyr,Thr

permease of ammonia

permease of ammonia

permease of ammonia









nutrient sensing system



Nitrogen catabolite repression regulator



Nitrogen catabolite repression regulator









The molecular and cellular biology of the yeast Saccaromyces: Gene expression (volume 2). Eds:

Substrate specificity and gene expression of the amino acid permeases in S. cerevisiae

Sensors of extracellular nutrients in S. cerevisiae

Nitrogen regulation in S. cerevisiae

Nitrogen catabolite repression in S. cerevisiae

Functional profiling of the S. cerevisiae genome

General and specific controls of lysine biosynthesis in S. cerevisiae

Gcn4p, a master regulator of gene expression, is controlled at multiple levels by diverse signals of starvation and stress.

A protein complex of translational regulators of GCN4 mRNA is the guanine nucleotide-exchange factor for translation initiation fact

Repression of GCN4 Translation by nitrogen starvation in S. Cerevisiae

In S. cerevisiae, feedback inhibition of homocitrate synthase by lysine modulate the activation of LYS gene expression by lysine

LYS80p of S. cerevisiae, previousely proposed as a specific repressor of LYS genes, is a pleiotropic regulatory factor identical to M

MKS1p is required for negative regulation of retrograde gene expression in S. cerevisiae but does not affect nitrogen catabolite repr

Control of enzyme synthesis in the lysine biosynthetic pathway of S. cerevisiae

Funtion Reference









-







-

Let aa_permease be the list of permeases of the amino acid.

Let Eaa be the external value of the amino acid.

Set aa_biosynthesis: if (INH3 >0) {aa_biosynthesis =1} -

else {aa_biosynthesis =0}.

if (max(aa_permeases) == 0) {retrun aa_biosynthesis}

else {return Eaa}



if (max(MEP1ap,MEP2ap,MEP3ap3) == 0) {return 0} -

else {return ENH3}

if (MKS1ap >=1) {return 0}

else if (min(LYS20,LYS21) ==0) {return 2} 1,12,13

else {return 1}

if (max(LYS20ap,LYS21ap)==0 OR IAlphaKetoGlutarate == 0) {return 0}

else: if(min(YJL200Cap,LYS2ap,LYS12ap,LYS4ap)==0) {return 2} 1

else {return 1}

if (min(YJL200Cap,LYS2ap,LYS1ap,LYS4ap) OR IHomoCytrate == 0) {return 0}

else: if(min(LYS9ap,LYS1ap)==0) {return 2} 1

else {return 1}

Set TranLys: if(LYP1ap==0) {TranLys=0} else {TranLys:=Elys}.

if (min(LYS9ap,LYS1ap)==0 OR IAlphaASAA == 0) {return TranLys} 1

else {return max(1, TranLys)}







1 -

1 -

1 -

1 -

Let min_Iaa=minimum value of all internal amino acids.

if (min(GCN2m,GCN1ap,GCN20ap) >=1) {return (2 - min_Iaa)} 8,9

else {return 0}

if (min(eIF3 subunits) == 0) {return 0}

else: if (INH3 == 0) {return GCN2ap - 1} 10,8,9

else {return GCN2ap}

if (LYS14m == 0) {return 0}

else: if (ASAA == 0} {return 1} 14

else: {return 2}

if(MKS1m == 0) {return 0} 12,13

else {return (2-INH3)}







1

1

1

1

1 9

1

1

1

1

1

equal to GCN3m

equal to GCD1m

equal to GCD6m

equal to GCD7m

equal to GCD2m

equal to GCD11m

equal to SUI2m

equal to SUI3m

equal to GCN20m

equal to GCN1m









LYS14ap+GCN4ap 14,7,1

1 1

1 1

1 1

LYS14ap+GCN4ap 14,7

1 1

LYS14ap+GCN4ap 14,7

LYS14ap+GCN4ap 14,7

1 1

equal to LYS1m 1

if(LYS5ap == 0) {return 0}

else {return LYS2m} 1

equal to LYS4m -

equal to LYS5m -

equal to LYS9m -

equal to LYS12m -

if (Ilys == 2) {return 0}

else {return LYS20m} 11

if (Ilys == 2) {return 0}

else {return LYS21m}

equal to YJL200Cm -







min(GLN3ap,GAT1ap) [NCR*]

2,4,5



1

1

1

1

1 2

1

1

1

1

1

Let min_Iaa=minimum value of all internal amino acids.

min(SSY1ap, GRR1ap,min_aa)



2,3





1

1 2

1



equal to GAP1m

equal to CAN1m

equal to PUT4m

equal to MUP3m

equal to DIP5m

equal to SAM3m

equal to AGP3m

equal to MUP1m

equal to HIP1m

equal to APL1m

equal to MMP1m 2

equal to AGP2m

equal to LYP1m

equal to AGP1m

equal to GNP1m

equal to TAT2m

equal to TAT1m

equal to BAP2m

equal to BAP3m

equal to MEP1m

equal to MEP2m

equal to MEP3m







1

1

1 3

1

equal to SSYm

equal to GRR1m

if(GLN3m==0){return 0}

else {return (2-min(IGln,Iglu))} 4,5

if(GAT1m==0){return 0}

else {return (2-min(IGln,Iglu))}







cold spring harbour laboratory press, Cold Spring Harbo

Curr. Genet 36: 317-328 (1999(

Curr genet 40:91-109 (2001)

gene 290: 1-18 (2002)

Molecular Biotechnology 12:35-73 (1999)

Nature 418: 387-391 (2002)

Curr. Genet. 9:341-344 (1985)

gnals of starvation and stress. Eukaryot Cell. 2002 Feb;1(1):22-32.

change factor for translation initiation factor 2 in yeast. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5350-4.

The journal of biological chemistry 276:25661-25671 (2

tion of LYS gene expression by lysine Eur. J. Biochem 261:163-170 (1999)

pleiotropic regulatory factor identical to Mks1p Yeast 13: 1337-1346 (1997)

but does not affect nitrogen catabolite repression-sensitive gene expression. The journal of biological chemistry 277:20477-20482 (2

Eur. J. Biochem 171: 171-176 (1988)

comments on function

determined by the medium









determined by the medium

amino acid may be transported

by its permease into the

cell, and biosynthesized.









NH3 enters the cell through

its permeases



MKS1down regulate CIT2 needed

to synthesis of IAlphaKetoGlutarate









Lysine is transported by its

permease (LYP1), and

biosynthesized.









INh3 effect MKS1 activity

General constrol of GCN4 and

lysine pathway-specific control

of LYS14.









feedback inhibition of homocitrate

synthase by lysine modulate

the activation









a simplification of

Nitrogen catabolite repression

a great simplification of the

nutrient sensing system









Nitrogen catabolite repression

triggered by presence of Glu and Gln









cold spring harbour laboratory press, Cold Spring Harbor, New-York, 1992.

Curr. Genet 36: 317-328 (1999(

Curr genet 40:91-109 (2001)

gene 290: 1-18 (2002)

Molecular Biotechnology 12:35-73 (1999)

Nature 418: 387-391 (2002)

Curr. Genet. 9:341-344 (1985)

Eukaryot Cell. 2002 Feb;1(1):22-32.

Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5350-4.

The journal of biological chemistry 276:25661-25671 (2001)

Eur. J. Biochem 261:163-170 (1999)

Yeast 13: 1337-1346 (1997)

The journal of biological chemistry 277:20477-20482 (2002)

Eur. J. Biochem 171: 171-176 (1988)

Variable Type Variable name Description

External conditions EOC External Osmotic Conditions

HOG pathway - Sln1 branch(negative regulator)

mRNA SLN1m

YPD1m

SSK1m

SSK2m

SSK22m

active protein SLN1ap * Osmosensor.









YPD1ap Histidine kinase. Negative regulator.

SSK1ap Aspartyl kinase



SSK2ap MAPKKK





SSK22ap MAPKKK - analogous to SSK2







HOG pathway - Sho1 branch (positive regulator)

mRNA SHO1m

CDC42m

CDC24m

STE20m

CLA4m

STE50m

STE11m



active Protein SHO1ap 4 transmembrane domains, SH3 domain.

Probably not an osmosensor by itself

CDC42ap Rho-like G-Protein. Binds Ste20 and Cla4

CDC24ap Exchange factor for Cdc42

STE20ap Kinase



CLA4ap similar function to Ste20

STE50ap Regulator.

Constitutively bound to Ste11 (ref. 13)

STE11ap MAPKKK



HOG pathway - general

mRNA PBS2m





HOG1m



PTP2m

PTP3m



PTC1m

PTC2m

PTC3m

active Protein PBS2ap MAPKK







Scaffold (in Sho1 branch)



cytHOG1ap MAPK

* Hog1 in Cytoplasm

* Active Hog1 is phosphorilated in both Tyr and

Thr residues





nuHOG1ap Hog1 in Nucleus









PTP2ap Tyrosine phosphatase

PTP3ap Tyrosine phosphatase

PTC1ap Serine/Threonine phosphatase

PTC2ap Serine/Threonine phosphatase

PTC3ap Serine/Threonine phosphatase



Hog1 translocation

mRNA GSP1m

NMD5m

active Protein GSP1ap

NMD5ap



TFs

Hot1m

Msn1m

Msn2m

Msn4m

active Protein Hot1ap * Recruits Hog1 to target promoters.

Most likely binds Hog1 in the nucleus.

* Has a physical interaction with Hog1

* Phos. upon osmotic sress - mainly by Hog1.

* Phosphorilation may be required for activity

Msn1ap Nuclear. Binding site similar to Hot1.

Most likely binds Hog1 in the nucleus

Does not need to be phosphorilated to be active.

Msn2ap Cyt+Nuc. Activator. Recognizes STRE.

Most likely binds Hog1 in the nucleus

Activated (phosphorilated) by PKA / Hog1

Msn4ap Cyt+Nuc. Activator. Recognizes STRE.

Most likely binds Hog1 in the nucleus

Activated (phosphorilated) by PKA / Hog1



PKA

mRNA PKA1m

PKA2m

PKA3m

active Protein PKAap



Osmotic-stress induced

mRNA GPD1m G3P dehydrogenase









GPP1m

GPP2m G3 phosphatase

GLO1m

DAK1m

TPK1m PKA1 (see above)

active Protein GPD1ap NAD1-dependent sn-glycerol 3-phosphate dehydrogenase

GPP1ap

GPP2ap

GLO1ap

DAK1ap

TPK1ap



Glycerol export channel

Fps1 * Glycerol transporter

* not part of the HOG pathway!





bibliography

1 S. Hohmann Osmotic Stress Signaling and Osmoadaptation in Yeasts

2 Unique and Redundant Roles for HOG MAPK Pathway Components as Revealed by W

S.M.O'Rourke and I. Herskowitz

3 Yeast go the whole HOG for the

S.M.O'Rourke, I. Herskowitz and E.K. O'Shea hyperosmotic response

4 The Transcriptional Response of Saccharomyces cerevisiae to Osmotic Shock

M. Rep, M. Krantz, J. M. Thevelein and S. Hohmann

5 A Third Osmosensing Branch in S.C. Requires the Msb2 protein and Functions in parall

S.M.O'Rourke and I. Herskowitz

6 Two protein-tyrosine phosphatases C, Ota I.

Jacoby T, Flanagan H, Faykin A, Seto AG, Mattisoninactivate the osmotic stress response pathway in ye

7 Ptc1, a type 2C Amin D, Ota I.

Warmka J, Hanneman J, Lee J,Ser/Thr phosphatase, inactivates the HOG pathway by dephosphorylatin

8 HOG on

Srikumar P. Chellappan the Promoter: Regulation of the Osmotic Stress Response

9 Regulation of the Saccharomyces

Wurgler-Murphy SM, Maeda T, Witten EA, Saito Hcerevisiae HOG1 mitogen-activated protein kinase by

10 Osmostress-induced transcription by Hot1

Alepuz PM, de Nadal E, Zapater M, Ammerer G, Posas F. depends on a Hog1-mediated recruitment of

11 A signaling mucin at the head of the Cdc42- and MAPK-dependent filamentous growth p

Cullen PJ, Sabbagh W Jr, Graham E, Irick MM, van Olden EK, Neal C, Delrow J, Bardwell L, Sprague G

12 Protein-protein interaction BJ, Davidson crucial role in controlling the sho1p-mediated s

Marles JA, Dahesh S, Haynes J, Andrews affinity plays aAR

13 Functional Characterization of the Interaction of Ste50p with Ste11p MAPKKK in Saccha

Cunle Wu, Ekkehard Leberer, David Y. Thomas, and Malcolm Whiteway

14 F, Saito H

Raitt DC, PosasYeast Cdc42 GTPase and Ste20 PAK-like kinase regulate Sho1-dependent activation o

15 MAP KinaseAlbertyn, Matthew Alexander, and Kenneth Davenport

Michael C. Gustin, Jacobus Pathways in the Yeast Saccharomyces cerevisiae

Funtion Reference

0 - normal, 1 - moderate stress, 2 - high stress



1

1

1

1

1

Sln1m AND (2 - EOC) 1









Ypd1m AND Sln1ap

Ssk1m AND Ypd1ap

9

Ssk2m AND Ssk1ap 1





Ssk22m AND Ssk1ap 1









1

1

1

1

1

1

1



Sln1m AND EOC



1

Cdc42m AND Cdc24 AND Sho1ap AND Pbs2ap?

CDC24m

Ste20m AND Cdc42ap 1



Cla4m AND Cdc42ap

Ste50m



13

Ste11m AND (Ste20ap OR Cla4ap) AND Ste50ap





1





1



max (1, nuHOG1ap) 5

6



max (1, nuHOG1ap) 6



1

1

1

Ssk2ap OR Ssk22ap OR 3

(Sho1ap AND Ste11ap)

7



1



Pbs2ap AND (NOT 3

(PTP2ap OR PTC1ap)) 1

1

6,7





(cytHOG1ap AND GSP1ap AND 1

NMD5ap) AND

(NOT (PTP2ap OR PTC1ap))

q 1

6,7

PTP2m 6

PTP3m 6

PTC1m 7

PTC2m 7

PTC3m 7





1

1

1 1

1 1





1

1

1

1

Hot1m AND Hog1ap 1



8





Msn1m 1



10

Msn2m AND (PKAap OR Hog1ap) 1



8

Msn4m AND (PKAap OR Hog1ap) 1



8





max(1, Hog1ap) 4

1

1

PKA1m AND PKA2m AND PKA3m





Hog1ap AND Hot1ap AND Msn1ap 4,8

8









1

Hog1ap AND Hot1ap 4

Hog1 AND (Msn2 OR Msn4) 4

Hog1 AND (Msn2 OR Msn4) 4

max(1, Hog1ap) 4

GPD1m

GPP1m

GPP2m

GLO1m

DAK1m

TPK1m





opposite of EOC 15









ptation in Yeasts

MAPK Pathway Components as Revealed by Whole-Genome Expression Analysis

motic response

aromyces cerevisiae to Osmotic Shock

quires the Msb2 protein and Functions in parallel with the Sho1 branch

ivate the osmotic stress response pathway in yeast by targeting the mitogen-activated protein kinase, Hog1.

activates the HOG pathway by dephosphorylating the mitogen-activated protein kinase Hog1.

Osmotic Stress Response

siae HOG1 mitogen-activated protein kinase by the PTP2 and PTP3 protein tyrosine phosphatases

1 depends on a Hog1-mediated recruitment of the RNA Pol II

42- and MAPK-dependent filamentous growth pathway in yeast

crucial role in controlling the sho1p-mediated signal transduction pathway in yeast

ction of Ste50p with Ste11p MAPKKK in Saccharomyces cerevisiae

e kinase regulate Sho1-dependent activation of the Hog1 MAPK pathway

haromyces cerevisiae

comments on function









Sln1 - Ypd1 - Ssk1

(phosphorelay system)

Histidine Kinase is activated by hypo-

osmolarity (cell swelling) and inhibited by

hyper-osmolarity (cell shrinking).

* Auto-phosphorilates itself under low

osmolarity

phosphorilated on His by Sln1

phosphorilated on Asp (+ stabilized) by Ypd1

(Phosphorilated form is inactive)

Activated by de-phosphorilated

Ssk1 (val should be opposite of Ssk1ap)

Activation involves auto-phosphorilation

Activated by de-phosphorilated

Ssk1 (val should be opposite of Ssk1ap)

Activation involves auto-phosphorilation









* Activated by high external osmolarity.

* Activation may involve demasking of SH3 domain

Activated by Sho1-Pbs2 complex (how?)



localization to the

cell surface + activation requires Cdc42



Didn't find evidence of change in activation/expression due to hyperosmitic conditions

In fact evidence shows it is constitutively bound to Ste11

Activated (phos.) by Ste20 (or Cla4). Regulated by Ste50









* Supposedly induced - didn't find evidence

* Induction may involve Msn2/4 as well as

other elements.





Transcription may be regulated by HOG pathway (how?)

Transcription may be regulated by HOG pathway (how?)

Transcription may be regulated by HOG pathway (how?)

* Phosphorilated and activated by MAPKKK

(Ssk2/22 or Ste11). Phos. On Ser and Thr.

* Supressed (de-phosphorilated and/or

blocked) by Ptc2/3

* In the Sho1 branch - recruited to the cell

surface by Sho1 binding in its SH3 domain (this is crucial)

* Phosphorilated and activated by MAPKK

Pbs2, may also be auto-phosphorilated

* Phos. On Thr and Tyr.

* Phosphorilated in the cytosol!

* Supressed (de-phosphorilated and/or

blocked) by Ptp2/(3) and Ptc1

* Activated (P) Hog1 is translocated into the nucleus

by Gsp1p and Nmd5p.

* Dephosphorylation and export from the nucleus are probably separated

* Supressed (de-phosphorilated and/or

blocked) by Ptp2/(3) and Ptc1

Expression induced by Hog1???









* Induced by Hog1, Hot1, Msn1

* Hot1 binds directly to promoter

* Increased binding upon osmotic stress

requires Hog1.

* Only phosphorilated Hog1 binds to promoter,

and binding requires Hot1.



Induced by Hog1, Hot1 (a lot less than the others)

Induced by Hog1, Hot1

Msn2/4 are redundant

Msn2/4 are redundant









inhibited by hyperosmotic stress in a manner

independent of the HOG pathway









-Genome Expression Analysis





ith the Sho1 branch

by targeting the mitogen-activated protein kinase, Hog1.

he mitogen-activated protein kinase Hog1.



PTP2 and PTP3 protein tyrosine phosphatases





al transduction pathway in yeast

myces cerevisiae

e Hog1 MAPK pathway



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