Molecular signatures of cold-adaptation in the

Conclusion Comparative se- quence analyses of zoarcid con- geners revealed patterns of a dif- ferent thermal adaptation at the molecular level although their mean habitat temperature differs only by ~10°C. The compositional

biases of cold-adapted proteins and DNA/RNA support an increa- se of molecule flexibility at the re- spective levels. These trends pro- mote optimized reaction kinetics at low temperatures trough less stable transition states in reaction

processes shaping the thermal plasticity of organisms at a deeper level. Finally, these data highlight another aspect of spe- cies‘ vulnerability of cold-adapted organisms in the context of the ongoing climate change.

Molecular signatures of cold-adaptation in the

Antarctic eelpout (Pachycara brachycephalum)

SSP Antarctic surveys, LU 1463/1-1

Biosciences

Integrative Ecophysiology Dr. Heidrun Windisch Am Handelshafen 12 27570 Bremerhaven fon +49-471-4831-2288 mail Heidrun.Windisch@awi.de web www.awi.de

YOUMARES | 4

Poster ID VII-1

Heidrun Windisch , Magnus Lucassen & Stephan Frickenhaus

Introduction Adaptation of ec- totherm organisms to the key abiotic factor temperature can occur by the development of ad- vantegous traits at different levels of organization [1]. Since all biological processes are sub- jected to the same physicoche- mical paradigms (Q10-rule), the

composition of biological macro- molecules is likely to be affected as well. Hence, proteins and nuc- leic acids may also contribute to a certain thermal plasticity and tolerance.

Antarctic fishes have adapted to constant cold environmental con- ditions for millions of years and

provide therefore excellent study objects to evaluate evolutionary trends of thermal adaptation.

Comparative sequence analyses with temperate species shed light on the molecular architecture of biological macromolecules in connection with species habitat temperatures.

Methods DNA/RNA sequences as well as their respective trans- lations were compared between the Antarctic eelpout Pachycara brachycehalum and its tempera- te congener, the Northsea Eel- pout, Zoarces viviparus through analyses of normalized transcriptomic cDNA libraries.

Zoarces viviparus Pachycara brachycephalum

reads total 481,802 ~400,000 in assembly 338,993 349,102

mean length 321 221

total 65,565 53,447

contigs reads

annotation 19,460 16,315

4,155 orthologs

known function matching

sequence pairs

Pachycara [2] Zoarces [3]

overview of cDNA libraries

Results

Purine metabolism

Secondary bile acid biosynthesis

metabolism Phenylalanine

Chloroalkane and cloroalkene degradation

isoleucine degradation Valine, leucine and metabolism

Glycerolipid

Gluconeogenesis Glycolysis /

tryptophan biosynthesis Phenylalanine , tyrosine and

metabolism Pyruvate

Taurine and hypotaurine metabolism Arachidonic acid

metabolism

Butanoate

metabolism Fatty acid elongation in mitochondria

biosynthesis Clavulanic acid

by cytochrom P450 Metabolism of xenobiotics

Selenocompound metabolism

biosynthesis Insect hormone

metabolism Sphingolipid

metabolism Methane [B] Phytochemical

compounds

degradation Naphthalene

polyketide products Biosynthesis of type II

isoleucine biosynthesis Valine, leucine and biosynthesis

Isoflavonoid

Fructose and mannose metabolism

resistance beta-Lactam [B] Proteoglycans

Brassinosteroid biosynthesis

Fatty acid metabolism

Glutathione metabolism

Tyrosine metabolism Glycosphingolipid

biosynthesis -

lact and neolacto series

metabolism Caffeine

Anthocyanin biosynthesis

hydrocarbon degradation Polycyclic aromatic

pyridine alkaloid biosynthesis Tropane, piperidine and biosynthesis

Carotenoid

C5-Branched dibasic acid metabolism

biosynthesis Indole alkaloid

metabolism Porphyrin and chlorophyll N-Glycan

biosynthesis

biosynthesis cephalosporin Penicillin and globoseries

Glycosphingolipid biosynthesis -

Inositol phosphate metabolism

biosynthesis Streptomycin

metabolism beta-Alanine unit biosynthesis

Polyketide sugar

Nitrogen metabolism

DDT degradation degradation

Benzoate

Glycine, serine and threonine metabolism degradation

Glycosaminoglycan

biosynthesis Lysine biosynthesis

Monoterpenoid

degradation Toluene

proline metabolism Arginine and metabolism

dicarboxylate Glyoxylate and

biosynthesis Novobiocin

biosynthesis Peptidoglycan

metabolism Riboflavin

biosynthesis Steroid hormone

Sulfur metabolism

by folate One carbon pool

proteins

[B] Lipids biosynthesis

Pentose and glucuronate interconversions

Biotin metabolism

other terpenoid-quinone biosynthesis

Ubiquinone and biosynthesis

Fatty acid

metabolism Pentose phosphate

degradation Xylene Terpenoid backbone

biosynthesis

degradation Bisphenol

D-glutamate metabolism D-Glutamine and Steroid biosynthesis

Biosynthesis of type II polyketide backbone

Aminobenzoate degradation

metabolism Vitamin B6

Dioxin degradation Biosynthesis of

unsaturated fatty acids

biosynthesis Puromycin

metabolism D-Alanine

metabolism Lipoic acid Cutin, suberine and wax biosynthesis

Nitrotoluene degradation [B] Glycosyltransferases

Butirosin and neomycin biosynthesis

ganglioseries Glycosphingolipid biosynthesis -

Glycosaminoglycan biosynthesis - chondroitin sulfate

Pantothenate and CoA biosynthesis

degradation of Ketone bodies Synthesis and

metabolism Thiamine

Ether lipid metabolism

Flavonoid biosynthesis

carboxylate cycle Reductive (CO2 fixation) Glycosaminoglycan heparan sulfate biosynthesis -

metabolism

Cystein and methionine

glutamate metabolism Alanine , aspartate and Other types of

O-glycan biosynthesis

Nicotinate and nicotinamide metabolism metabolism

Amino sugar and nucleotide sugar

biosynthesis Zeatin

Ascorbate and aldarate metabolism

pinene degradation Limonene and

metabolism Propanoate

degradation Caprolactam

N-glycan biosynthesis Various types of

Phenylpropanoid biosynthesis

phosphorylation Oxidative

Histidine metabolism

metabolism Tryptophan

Citrate cycle (TCA cycle) biosynthesis

Sesquiterpenoid

Other glycan degradation

Mucin type

O-glycan biosynthesis

ansamycins Biosynthesis of

metabolism Retinol

metabolism Glycerophospholipid

biosynthesis Primary bile acid

Photosynthesis - antenna proteins

D-ornithine metabolism D-Arginine and

biosynthesis Glucosinolate biosynthesis

Flavone and flavonol

[B] Cytochrome P450

biosynthesis Folate

Photosynthesis

Pyrimidine metabolism metabolism

alpha-Linolenic acid biosynthesis -

keratan sulfate Glycosaminoglycan

biosynthesis Lipopolysaccharide

Galactose metabolism

[B] Lipids

fixation Carbon

biosynthesis Isoquinoline alkaloid

nonribosomal peptides siderophore group Biosynthesis of

Chlorocyclohexane and chlorobenzene degradation Phosphonate and

phosphinate metabolism

Tetracycline biosynthesis degradation

Atrazine

degradation Styrene

biosynthesis Benzoxazinone metabolism

Starch and sucrose

- other enzymes Drug metabolism

(GPI)-anchor biosynthesis Glycosylphosphatidylinositol

metabolism Linoleic acid

16-membered macrolides Biosynthesis of 12-, 14- and

Ethylbenzene degradation

proteins

[B] Photosynthesis

Cyanoamino acid metabolism

Biosynthesis of vancomycin group antibiotics

- cytochrom P450 Drug metabolism degradation

Fluorobenzoate

Lysine degradation

biosynthesis Diterpenoid

biosynthesis Acridone alkaloid

Metabolism of

Terpenoids and Polyketides

Nucleotide Metabolism and Metabolism

Glycan Biosynthesis

Amino Acid Metabolism

Metabolism of Other Amino Acid and Metabolism

Xenobiotics Biodegradation

Other Secondary Metabolites Biosynthesis of

Metabolism Energy Carbohydrate

Metabolism

Cofactors and Vitamins Metabolism of

Metabolism Lipid

Purine m

etabo

lism

Seconda ry bil

e ac

id

biosynthesis metabolism

Phenylala

nine

Chloro alkane and

cloro

alk

ene de gradati on isoleucine degra

datio

n Valine, leucine and

metab

olismGlyce

rolipid

Gluconeog enes

is Glycolysis /

tryptop han bi osynthes

is Phenylalanine , tyrosine and

metabolism Pyruvate

Tauri

ne and hypotaurine metab olism

Arachid

onic ac

id

meta bolism

Butano

ate meta bolism Fatty acid elo ngatio

n

in m itochondria biosynthe

sis Clavulanic ac

id

by cytoc

hrom P450 Metabolis

m of xenob iotics Selenoc

ompo

und metab

olis

m

biosynthesis Insect hormone metab

olism Sphingol

ipid

metabolism ane Meth

[B] Phytochemical compounds

degrad

ation Naphthal

ene

poly

keti

de prod

ucts of tysisyntheBios

pe II

isoleuc

ine bi

osynthes

is Valine, leucine and

biosyn

thes

is Isoflavonoi

d

Fructose and mannose me tabol ism

resistanc

e beta-Lactam

[B] Proteoglycans

Bra

ssinos

teroi

d

biosynthesis Fatty acid

metab olism

Glutathione metab olism

Tyrosi

ne metabolism

Glycosphing

olip

id

biosynthe

sis - lact and neol

acto series

metab

olismCaffei

ne

Anthocyanin biosynthe

sis

hydrocarbon degrada

tionticromaic ayclPolyc

pyrid

ine a

lkal

oid b

iosyn

thesi

s Tropane, piperidine and

bios

ynthe

sis Carotenoid

C5-Branched dibasi

c ac

id

metabo lism biosyn

thesi

s Indole alka

loid

metab olism

Porphyrin and

chlo

rophyl

l N-Glycan

biosynth

esis

biosyn

thes

is cephalospori

n Penicillin and

globo serie

s

Glycosphing

olip

id biosynthesis -

Inositol phos

phate metabolism

biosynth

esis Streptomy

cin

metab

olism beta-A

lani

ne

unit

biosynthesis Polyketide

sugar

Nitrogen

meta bolism

DDT d egradation

degrada

tion Benzoate

Glycine, se rine an

d

threonine metabolism

degradati

on Glycosami

noglyca

n

biosynthesis

Lys

ine

biosynthe

sis Monoterpenoi

d

degradati

on eneTolu

proli

ne m

etabol

ism Arginine and

metabolis

mylateylatdicarboxGlyox

e and

biosyn

thes

is Novobiocin

bios

ynthe

sis Peptidogly

can

metab

olism Ribofl

avin

bios

ynthe

sis Steroid ho

rmone Sulfur

metabolis m

by

folate One carbon p

ool

proteins [B] Lip

ids

bios

ynthe

sis Pentose and glucuronat

e

interconversions

Bioti

nmetab

olism

other t

erpenoi d-quin one

biosyn thesi

s

Ubiquino

ne and id biosynthesis Fatty ac

metab

olism Pentos

e phos phate

degradati

oneXylen backbone oid Terpen

biosyn thesi

s

degradati

onhenolBisp

D-glutam ate

metabolis m

D-Glutam

ine a

nd

Steroid biosynthes

is

Biosynthesis of type II poly

keti

de bac

kbone

Aminobe

nzo

ate degradati on metab

olism Vitamin

B6

Dioxin degrad ation

Biosynthes is of

unsaturated fatty

ac

ids biosyn

thes

is Puromycin

metab olism D-Alanine

metab

olis

midc acLipoi

Cutin, sube

rine a

nd wax biosynth esis

Nitrotoluenedegradation

[B] Glycosyltransferases

Butiro

sin

and neomycin biosynthesis

gangl

iose

ries Glyco

sphi

ngol

ipid biosyn

is -thes

Glycosami nogly

can biosynthesis - chondroitin sulfate

Pantothe nate and CoA

biosyn thesi

s

degrada

tion of Ketone bod

ies Synthes is and meta

bolism Thiam

ine

Ether lipid metabol

ism

Flavonoid

biosynthe

sis

carbox

ylat

e cy

cle Reductive

(CO2 fi

xati

on) Glyco

sam

inogl

yca

n

heparan

sul

fateis -thes biosyn

metab

olismCystei

n and methio

nine

glutamate m etabol

ism Alanine , aspartate and

Other types of O-glyc

an bi

osynthes

is

Nico

tinate

and

nicotinamide metab olism

meta bolism

Am

ino su

gar and nucleoti de su gar

biosynthesis Zeatin Ascorbate and

ald

arate me tabol ism

pine

ne deg radatio

n Limonene and

meta

bolismpanoate Pro

degrada

tion Caprolactam

N-glyc

an bi

osynthes

is Various types of

Phenyl

propan

oidsthesi biosyn

phos phoryl

ationve Oxidati

Histid ine metabol ism meta

bolismptophanTry

Citrate cy

cle

(TCA cy cle)

biosyn thesi

s Sesquiterpeno

id

Other g

lycan degrada

tion

Mucin

type

O-glycan bi

osy

nthes

is

ansamycins Biosynthesis of

metab

olism Retinol

metabolis

m Glyceropho

sphol

ipid biosynthesis Primary bile acid

Photos

ynthe

sis -

antenn a protei ns

D-ornithin

e

metab olism

D-Argin

ine a

nd

bio

synthesis Glucosi

nol

ate

biosyn thesi

s Flavone and

flavonol

[B] Cytoch rome P450 biosynthesis

Folat

e

Photos

ynthe

sis

Pyrimidine metabol

ism metabolism alpha-Linolenic acid

biosyn thesi

s -

kerat an sulfa te

Gly

cos

ami

noglycan biosynthe

sis Lipopolys

accharide

Galactos

e

metab olism

[B] Lipids

fixa

tionCarbon

biosynthe

sis Isoquinoline

alkal

oid

nonrib osomal

peptides

side

rophore

groupis of Biosynthes

Chl

orocyclohexane and chlorobe

nze

ne degradation Phosphonat e and

phosphinate metabo lism

Tetracyc

line

biosynthesis

degradation

Atrazi

ne

degradat

ion Styrene

biosynthes

is Benzoxazinone

metab

olismStarch an

d su

cros

e

- other enzymes

Dru

g met abolism (GPI)-anc hor biosyn

inosthesisdyl lphosphati Glycosy

itol

metabolism

Lin

oleic acid 16-mem

bered macrol

ides sis of 12-, 1 yntheBios

4- and

Ethylbe

nze

ne degrada tion proteins

[B] Photos

ynthe

sis

Cyanoamino a

cid

metab

olism

Biosy

nthes

is of va

ncomyci

n

group an tibioti cs

- cyto

chrom

P450 Drug metabolis m

degradati

onenzoa Fluorob

te

Lys

ine

degradati on

biosynthe

sis Diterpenoid

biosynthe

sis Acridone

alkaloi

d

Metabolism of Ter penoids and Polyketides Nucleotide

Metabolism and Metabolism Glycan Biosynthesis

Amino Acid Metabolism

Metabolis

m of Other Amino

Aci d

and Metabolism Xenobiotics Biodegradation Other Secondary Metabolites

Biosynthesis of

Metabolism Energy

Carbohydrate Metabolism Cofactor

s and Vitamins Metabolism of

Metabolism Lipid

Functional differences

Translation, ribosomal structure and biogenesis RNA processing and modification Transcription Replication, recombination and repair Chromatin structure and dynamics Cell cycle control, cell division, chromosome partitioning Nuclear structure Defense mechanisms Signal transduction mechanisms Cell wall/membrane/envelope biogenesis Cell motility Cytoskeleton Extracellular structures Intracellular trafficking, secretion, and vesicular transport Posttranslational modification, protein turnover, chaperones Energy production and conversion Carbohydrate transport and metabolism Amino acid transport and metabolism Nucleotide transport and metabolism Coenzyme transport and metabolism Lipid transport and metabolism Inorganic ion transport and metabolism Secondary metabolites biosynthesis, transport and catabolism General function prediction only Function unknown

G. aculeatus Z. viviparus

P. brachycephalum

0 1,000 2,000 3,000 4,000 5,000

counts meNOGs S

R Q P I H F E G C O U W Z N M T V Y D B L K A J

Functions, differently represented in respective transcripto- mes may indicate gene duplication events and distinct meta- bolic demands. As a „genomic scale“, the functional distributi- on of Gasterosteus aculeatus is indicated by green bars.

Amino acid usage

0 200 400 600 800 1,000

Z. viviparus

P. brachycephalum

1,200 total counts of amino acids in non-synonymous sequence pairs

F G K

L M N

P Q R

S T

W Y A

unpolar, hydrophobic V E D

H

C

I

polar, neutral acidic basic

Imbalances in the amino acid usage are pronounced by 102 fewer ionic aa in P. brachycephalum reducing ionic interac- tions of proteins. A gain of 117 polar/neutral aa increases pro- tein polarity as well as better solvent-interaction. However, no clear pattern exists within the unpolar/hydrophobic group.

Codon usage

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