Thesis report on Genetic study of Bhumij Tribe of Jharkhand using mt DNA and Y chromosome DNA marker by Smita Bernadet Kujur - HTML preview

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Introduction To The Study

INTRODUCTION:

Tracing about the origin and ancestral links of homo sapiens have

been the subject of curiosity for various scientists. And a number of

scholars have devoted themselves to disclose these hidden mysteries

of Human origin and dispersal on earth.

Where did we come from, and how did we get here? This is the

question which genetic anthropology field is seeking an answer for.

DNA studies indicate that all modern humans share a common

female ancestor who lived in Africa about 140,000 years ago, and all

men share a common male ancestor who lived in Africa about 60,000

years ago. These were not the only humans who lived in these eras,

and the human genome still contains many genetic traits of their

contemporaries. Humanity’s most recent common ancestors are

identifiable because their lineages have survived by chance in the

special pieces of DNA that are passed down the gender lines nearly

unaltered from one generation to the next. These ancestors are part of

a growing body of fossil and DNA evidence indicating that modern

humans arose in sub-Saharan Africa and began migrating, starting

about 65,000 years ago, to populate first southern Asia, China, Java,

and later Europe. Each of us living today has DNA that contains the

story of our ancient ancestors’ journeys.

When DNA is passed to our next generation, the processes that make

each person unique from their parents is the combination of both their

genomes. Some special pieces of DNA, however, remain virtually

unaltered as they pass from parent to offsprings. One of these pieces

are carried by Y chromosome. It is passed only from father to son.

Secondly, mitochondrial DNA (mtDNA), is passed (with few

exceptions) only from mother to child. Since the DNA in the Y

chromosome does not undergo crossing over, it is like a genetic

surname that allows scientists to trace back their paternal lineages.

Similarly, mtDNA allows both men and women to trace their

maternal lineages. Both the Y chromosome DNA and mtDNA are

subject to occasional harmless mutations that become inheritable

genetic markers. After several generations, almost all male and female

inhabitants of the region in which it arose carry a particular genetic

marker. When people leave that region, they carry the marker with

them. By studying the genes of many different indigenous

populations, scientists can trace when and where a particular marker

arose. Each marker contained in a person’s DNA represents a location

and migration pattern of that person’s ancient ancestors. For example,

roughly 70% of English men, 95% of Spanish men, and 95% of Irish

men have a distinctive Y-chromosome mutation known as M173. The

distribution of people with this mutation, in conjunction with other

DNA analysis, indicates that they moved north out of Spain into

England and Ireland at the end of the last ice age

(genomics.energy.gov).

Information about the history of our species comes from two main

sources: the paleo-anthropological record and historical inferences

based on current genetic differences observed in humans. Although

both sources of information are fragmentary, they have been

converging in recent years on the same general story (Underhill et;

al.).

Since the 1990s, it has become common to use multilocus genotypes

to distinguish different human groups and to allocate individuals to

groups (Bamshad et al. 2004). These data have led to an examination

of the biological validity of races as evolutionary lineages and the

description of races in cladistic terms. The technique of multilocus

genotyping has been used to determine patterns of human

demographic history. Thus, the concept of “race” afforded by these

techniques is synonymous with ancestry broadly understood (Berg et

al.,).

Y chromosome and mitochondrial DNA are transmitted uni-parentally

through father and mother, respectively and don’t under go any

recombination. Hence, markers present on both are useful to trace the

paternal and maternal lineages. Haplotypes can be constructed by

combining the allelic status of multiple markers, which would provide

adequate information for establishing paternal lineages. The non-

coding region (D-loop) of mtDNA, which harbors two hyper variable

regions (HVR I and HVRII), shows variation between different

populations. A large number of studies have been conducted on

various populations using Y chromosome markers and mtDNA D-

loop region to understand their origin, evolution and migration.

Indian populations reveal striking diversities in terms of language,

marriage practices as well as in their genetic architecture. The social

structure of the Indian population is governed by the hierarchical caste

system. In India, there are nearly 5,000 well-defined endogamous

populations. In addition to the native populations, there are a few

migrant populations inhabiting various parts of India. Several

important historical migrations into India caused amalgamation of

migrant populations with the local population groups. Major

demographic event like migrations, population bottlenecks and

population expansion leave genetic imprints and alter gene

frequencies. These imprints are passed onto successive generations,

thus preserving the population’s history within the population.

Therefore, we have undertaken to disclose the genetic information

about how different caste and tribal populations of India help to

construct ecognize and help to construct the evolutionary tree

(Cavalli-Sforza et al.,).

Two major routes have been proposed for the initial peopling of East

Asia; one via Central Asia to Northeast Asia, which subsequently

expanded towards Southeast Asia and beyond, and the other through

India to Southeast Asia and further to different regions of East

Asia.[1] It is pertinent in this context that the Indian subcontinent has

been considered as a major corridor for the migration of human

populations to East Asia.[2-4] Given its unique geographic position,

Northeast India is the only region which currently forms a land

bridge between the Indian subcontinent and Southeast Asia, hence

hypothesized as an important passage for the initial peopling of East

Asia. This region is inhabited by populations belonging to Indo-

European, Tibeto-Burman and Austro-Asiatic linguistic families.

‘‘BHUMIJ TRIBE’’ come under austro-asiatic linguistic

population. Austro-Asiatic speakers, hypothesized as probably the

earliest settlers in the Indian subcontinent ([5] and references their

in), are also found in other parts of India as well as in East/Southeast

Asia. Therefore, if Northeast India had served as an initial corridor, it

is likely that the Austro-Asiatic tribes of this region should provide

hitherto missing genetic link, which may reflect genetic continuity

between Indian and East/Southeast Asian populations. Based on

mitochondrial DNA (mtDNA) and Y-chromosome markers, Cordaux

et al. [6] observed genetic discontinuity between the Indian and

southeast Asian populations and inferred that Northeast India might

have acted as a barrier rather than the facilitator of the movement of

populations both into and out of India.

However, this study include only ‘‘BHUMIJ’’ Tribe of Jharkhand

region from Jamshedpur district. Further evidence is needed by way

of determining the mtDNA and Y-chromosome haplogroups/lineages

of the Austro-Asiatic tribes of the northeastern region and their

comparison with appropriate set of South and Southeast Asian

populations. Jharkhand is basically an agricultural land.

Geographically it is covered by jungles, mountains, rivers and

Chotanagpur plateau etc.

1.2 BACKGROUND :

HUMAN GENOME DIVERSITY PROJECT (HGDP) :

The HGD Project was started internationally on mid-September

of 1993 and it has 13 countries participating in it. The Human

Genome Diversity Project is an international project that seeks to

understand the diversity and unity of the entire human species.

The Human Genome Diversity Project (HGDP) aims to collect

biological samples from different population groups throughout the

world, with the aim of building up a representative database of human

genetic diversity. This seems a laudable aim, but the Project has been

enmeshed in massive controversy since it was first proposed in 1991,

with violent reactions from many of the indigenous people’s groups it

proposes to study.

The eminent geneticist Luigi Luca Cavalli-Sforza of Stanford

University first conceived by the HGDP. For many years, he and other geneticists and anthropologists have been visiting different ethnic

groups around the world, collecting samples, and trying to build up a

picture of how different human populations are related to each other.

The samples are seen as immensely valuable, but they are in

laboratories spread around the world. In 1991, Cavalli-Sforza and a

number of colleagues wrote a letter to the scientific journal, Genomics,

pointing out the need for a systematic study of the whole range of

human genetic diversity, within the context of the Human Genome

Project. They pointed to a problem: ‘The populations that can tell us

most about our evolutionary past are those that have been isolated for

some time, are likely to be linguistically and culturally distinct and are

often surrounded by geographic barriers. Such isolated populations are

being rapidly merged with their neighbours, however, destroying

irrevocably the information needed to reconstruct our evolutionary

history. It would be tragically ironic if, during the same decade that

biological tools for understanding our species were created, major

opportunities for applying them were squandered.

Major demographic events like migration, population

bottlenecks and population expansion leave genetic imprints where

gene frequency of the genome is altered (Thangaraj et, al., 1998).

These imprints are passed onto successive generations thus preserving

the population history within the population. In general, human

beings group themselves into units in such a way that members

between units rarely exchange genes due to cultural and

geographical barriers resulting in genetic divergence of population.

The Human Genome Diversity Project proposed in early nineties is a

combined effort preceded by anthropologists, geneticists, doctors,

linguists and other scholars from around the world aims at collecting

the blood samples from different ethnic populations throughout the

world aiming at building up a representative database of human genetic

diversity.

The reason lying behind selecting only tribes for sampling is that they

are believed to have been isolated during an evolutionary time,

linguistically and culturally distinct and are often isolated by

geographic barriers and thus prove to be best tools for study.

IN THIS PROJECT, THE SUBJECT OF GENETIC STUDY IS

‘‘BHUMIJ TRIBE’’ FROM JHARKHAND (CHOTANAGPUR

PLATEAU), INDIA .

1.3 STATEMENT OF PURPOSE :

How does DNA helps us to trace back?

Y chromosome and mitochondrial DNA are transmitted uni-parentally through father and mother respectively and do not undergo any

recombination. Hence, markers present on both are useful to trace the

paternal and maternal lineages. Haplotypes can be constructed by

combining the allelic status of multiple markers, which would provide

adequate information for establishing paternal lineages. The non-coding

region (D-loop) of mtDNA, which harbors two hyper variable regions

(HVR I and HVRII), shows variation between different populations. A

large number of studies have been conducted on various populations

using Y chromosome markers and mtDNA D-loop region to understand

their origin, evolution and migration.

Indian populations reveal striking diversities in terms of

language, marriage practices as well as in their genetic architecture. The

social structure of the Indian population is governed by the hierarchical

caste system. In India, there are nearly 5,000 well-defined endogamous

populations. In addition to the native populations, there are a few

migrant populations inhabiting various parts of India Several important

historical migrations into India caused amalgamation of migrant

populations with the local population groups. Major demographic event

like migrations, population bottlenecks and population expansion leave

genetic imprints and alter gene frequencies. These imprints are passed

onto successive generations, thus preserving the population’s history

within the population. Therefore, we have undertaken to disclose the

genetic information about caste and tribal populations of India to

construct ecognize and to use the ecognize data to construct the

phylogenetic tree.

In future the recorded data of mutated sites of a particular

haplogroup can help the scientists to trace the cause and solution to

many new diseases and help them to develop ne techniques of

diagnosis and design new drugs.

1.4 AIMS AND OBJECTIVES OF THE STUDY :

GOALS OF HGD PROJECT:

The Human Genome Diversity Project is a collaborative research project

that is being developed on a global basis under the auspices of the

Human Genome Organization (HUGO).

¾ The overall goal of the project is to arrive at a much more precise

definition of the origins of different world populations by integrating

genetic knowledge, derived by applying the new techniques for studying

genes, with knowledge of history, anthropology and language.

¾ To investigate the variation occurring in the human genome by

studying samples collected from populations that are representative of all

of the world’s peoples.

¾ To create a resource for the benefit of all humanity and for the

scientific community worldwide.

The resource will exist as a collection of biological samples that

represents the genetic variation in human populations worldwide and

also as an open, long-term, genetic and statistical database on variation

in the human species that will accumulate as the biological samples are

studied by scientists from around the world.

The major goals of HGDP:

¾ To identify all the approx 20,000-25,000 genes in human DNA,

determination of the sequence of the 3 billion chemical base pair that

make up human DNA.

¾ In silico storage of all DNA database. Improve tools for data analysis.

¾ Transfer related technologies to the private sector. Address the

ethical, legal and social issues (ELSI) that may arise from the project.

¾ To provide information regarding human biological relationship

among different groups and human history.

¾ To understand the cause and diagnostics of human diseases.

BENEFITS AND IMPLIFICATIONS OF HGDP:

The project will reap fantastic benefits for human kind, some that we can

anticipate and other that will surprise us. Generations of biologists and

researchers have been provided with detailed DNA information that will

be the key to understanding the structure, organization and function of

DNA in chromosome. The information from HGDP provides

information to clarify the origin and biological relationship of specific

human populations and the evolution of human being in particular. The

variations of frequencies in various populations can reveal how recently

they shared a large pool of common ancestors.

HGDP IN INDIA:

In India, Centre for Cellular and Molecular Biology [ CCMB], Hyderabad has pioneered the Human Genome Diversity Project in

collaboration with several other institutes and universities. Around

6,200 different unrelated individuals have been sampled from various

Indian populations & have been analyzed for their genetic diversity and

phylogeny.

The origins of Indian tribes, who presently constitute about 8% of total

population of India, have been subject to numerous genetic studies. India

is a land of enormous human genetic, bio-geographic, socio-economic,

cultural and linguistic diversity. More than 300 tribal groups are

recognized in India and they are densest in the central and southern

province. There are more than 800 dialects and a dozen major languages,

grouped into those of Dravidian South India and Indo-Aryan North

India. The resulting hypotheses range from referring to some tribes as

the descendents of the original Paleolithic inhabitants of India while

some are the recent immigrants. Hence, genetic diversity in India

provides important clues to the evolutionary history of human beings.

TRACING GENETIC DIVERSITY:

The past decade of advances in molecular genetic technology has

heralded a new era for all evolutionary studies, but especially the science

of human evolution. Data on various kinds of DNA variation in human

populations have rapidly accumulated. There is increasing recognition of

the importance of this variation for medicine and developmental biology

and for understanding the history of our species. Haploid markers from

mitochondrial DNA and the Y chromosome have proven invaluable for

generating a standard model for evolution of modern humans.

Conclusions from earlier research on protein polymorphisms have been

generally supported by more sophisticated DNA analysis. Co-evolution

of genes with language and some slowly evolving cultural traits, together

with the genetic evolution of commensals and parasites that have

accompanied modern humans in their expansion from Africa to the other

continents, supports and supplements the standard model of genetic

evolution. The advances in our understanding of the evolutionary history

of humans attest to the advantages of multidisciplinary research.

Although molecular genetic evidence continues to accumulate that is

consistent with a recent common African ancestry of modern humans, its

ability to illuminate regional histories remains incomplete. A set of

unique event polymorphisms associated with the non-recombining

portion of the Y-chromosome (NRY) addresses this issue by providing

evidence concerning successful migrations originating from Africa,

which can be interpreted as subsequent colonization, differentiations and

migrations overlaid upon previous population ranges. A total of 205

markers identified by denaturing high performance liquid

chromatography (DHPLC), together with 13 taken from the literature,

are used to construct a parsimonious genealogy. Ancestral allelic states

were deduced from orthologous great ape sequences. A total of 131

unique ecognize are defined which trace the micro evolutionary

trajectory of global modern human genetic diversification. The

genealogy provides a detailed phylogeographic portrait of contemporary

global population structure that is emblematic of human origins,

divergence and population history that is consistent with climatic, paleo-

anthropological and other genetic knowledge. The frequency of

occurrence of different ecognize can be used to distinguish

populations and to shed light on the sub-structures within a population,

also for inter and intra population variation studies. Population analyses

have examined allele frequencies at autosomal genetic markers (Cavalli-

Sforza As in this project, when a significant number of individuals in a

population et al., 1994). The incorporation of mitochondrial DNA during

the 1980s added a powerful tool to the geneticists’ tool kit, since mtDNA

does not recombine and is transmitted only through female germ line

(Stoneking and Soodyall, 1996). The increasing number of polymorphic

markers identified on the Y chromosome has allowed analyzing male

lineages, (Hammer and Zegura, 1997). A set of highly polymorphic

chromosome Y specific micro satellite became available for forensic,

population genetic and evolutionary studies. However, the lack of a

mutation frequency estimate for these loci prevents a reliable application.

MARKERS:

The human genome comprise of actually two genomes: a complex nuclear

genome, which account for 99.9995% of total genetic information and a

simple mitochondrial genome, which accounts for the remaining 0.0005%.

During zygote formation, a sperm cell contributes its nuclear genome, but

not its mitochondrial genome to the egg cell. Consequently, the

mitochondrial genome of the zygote is determined exclusively by that

originally found in the unfertilized egg. The mitochondrial genome is

therefore maternally inherited. As a result, it does not undergo any genetic

reshuffling and thus, is intact which makes it a unique tool for studying

human origins. Thus, everyone carries with them a more or less exact copy

of the mtDNA from their mother and their mother’s mother and so forth

for countless generations. The term “more or less exact” is the key to

scientist solving the mystery of human origins. That’s because like all

DNA, mtDNA is subject to random mutations over the eons. As these

mutations are passed on intact to next generation, they in effect become

“tracers” of family. A single type of circular double stranded molecule of

16,569 bases defines human mitochondrial genome.

MITOCHONDRIAL DNA (mtDNA) AS MARKER:

The mtDNA (Fig:1) has no repetitive DNA, spacers or introns. The

mtDNA contains 37 genes, all of which are involved in the production of

energy and its storage in ATP. It encodes 13 mRNAs, 22 tRNAs and 2

rRNAs. mtDNA has two strands, a guanine rich heavy (H) strand and a

cytosine rich light (L) strand. The heavy strand contains 12 of the 13-

polypeptide encoding genes, 14 of the 22 tRNA encoding genes and both